CZ129694A3 - Novel peptide derivatives - Google Patents

Abstract

The invention relates to new competitive inhibitors of thrombin, their synthesis, pharmaceutical compositions containing the compounds as active ingredients, and the use of the compounds as anticoagulants for prophylaxis and treatment of thromboembolic diseases, according to formula (I) wherein A represents a methylene group, an ethylene group or a propylene group, which may be substituted or A represents -CH2-O-CH2-, -CH2-S-CH2-, -CH2-SO-CH2-, or A represents -CH2-O-, -CH2-S-, -CH2-SO-, with the heteroatom functionality in position 4, or n is an integer 2 to 6; and B represents -N(R<6>)-C(NH)-NH2, wherein R<6> is H or a methyl group, or B represents -S-C(NH)-NH2, or -C(NH)-NH2. Further described is new use in synthesis of pharmaceutical compounds of a compound of formula (II).

Description

Technical field

The present invention relates to novel competitive thrombin inhibitors, to a process for their synthesis, to pharmaceutical compositions containing these compounds as active ingredients and to the use of these compounds as anticoagulants for the prophylaxis and treatment of thromboembolic diseases such as vein thrombosis, pulmonary embolism, arterial thrombosis. thrombosis, general hypercoagulable conditions, and local hypercoagulable conditions, for example after angioplasty as well as coronary bypass surgery.

The present invention also relates to a novel use of the compounds as starting materials for the synthesis of a serine protease inhibitor. The invention further relates to a novel structural fragment in a serine protease inhibitor.

BACKGROUND OF THE INVENTION

Blood coagulation is a key process that involves both haemostasis (that is, preventing blood loss from a damaged vessel) and thrombosis (that is, a pathological closure of a blood vessel by a blood clot). Coagulation is the result of a complex series of enzymatic reactions, where one of the final steps is the conversion of the prothrombin protease enzyme to the active enzyme thrombin.

Thrombin plays a central role in coagulation. It activates platelets, converts fibrinogen to monomeric fibrin, which spontaneously polymerizes into fibers, and activates coagulation factor XIII, which in turn crosslinks the polymer to non-degraded fibrin. Furthermore, thrombin activates factor V and factor VIII in a positive feedback reaction. Therefore, thrombin inhibitors are expected to be potent anticoagulants by inhibiting platelets, fibrin formation, and fibrin stabilization. By inhibiting the positive feedback mechanism, the agent is expected to induce an initially in-chain inhibition in a case that leads to coagulation and thrombosis.

This prior art is known.

Thrombin inhibitors based on the amino acid sequence around the fibrinogen Aa chain cleavage site were first described by Blomback et al. in J. Clin. Lab. Invest. 24, suppl.

107, 59/1969 /, which recorded on a the sequence Phe-Val-Arg _(P9 - ^P l ^-P 2 'which is here referred to as the ^{P3-P2-P 1-Vence)} to be the best inhibitor.

In U.S. Patent No. 4,346,078 (Richter Gedeon Vegyeszeti Gyar RT, priority date dated October 7, 1980) and S. Bajusz et al. in Peptides, pp. 643-347, Walter de Gruyter & (Berlin 1983) disclose a thrombin inhibitor H-DPhe-Pro-Agm, a dipeptidyl derivative with an aminoalkylguanine at the P 1 position.

S. Bajusz et al. in J. Med. Chem. 33, 1729-1735 (1990) and Richter Gedeon Vegyeszeti Gyar RT in EP-A2-0 185 390 (priority date 21 December 1984) also state that a replacement of agmatine argininaldehyde yields a thrombin inhibitor which has a much higher effect.

It is believed that the cause of the increased effect of this thrombin inhibitor is the possibility that hemiacetal is formed due to the interaction of the aldehyde function with the Ser-OH at the active site of the enzyme. It is not possible to have the same type of interaction in the dipeptide derivative H-DPhe-Pro-Agm because it does not contain an amino acid derivative with a carbonyl group in position.

Among other thrombin inhibitor work, serine proteinase inhibitors that are based on electrophilic ketones instead of the aldehydes at the P 1 position are included in the following publications:

Ε. N. Shaw et al. (Research Corporation), US Patent No. 4,318,904 (priority date April 25, 1980), describing a peptide of chloromethyl ketones, for example H-DPhe-Pro-Arg-CH ₂ Cl.

M. Szelke and DM Jones in EP-A1-0 118 280 (priority date 4 March 1983) describing compounds derived from the P ₂ - P ₂ 'pentapeptide sequence of the fibrinogen Aa chain in which the P ₂ - P ₁ ' peptide is readily cleavable the bond is replaced by a -CO-CH ₂ - residue that forms the keto isoster to the corresponding peptides.

M. Kolb et al. (Merrell-Dow), EP-A2-0 195 212 (priority date 4 February 1985), describing peptidyl-α-ketoesters and peptidyl-α-ketoamides.

B. Imperiali and R. H. Abes, Biochemistry 25, 3760 (1986), describing peptidylfluoroalkyl ketones.

D. Schirlin et al. (Merrell-Dow) EP-A1-0 362 002 (priority date 1 September 1988) describing fluoroalkyl amide ketones.

P. Bey et al. (Merrell-Dow), EP-A2-0 364 344 (priority date 1 September 1988) describing α, β, S-triketone compounds.

Ueda et al., Biochem. J. 265, 539 (1990), describing peptidyl fluoroalkyl ketones.

Thrombin inhibitors based on C-terminal orthoboric acid derivatives derived from arginine and isothiouronium analogs are described by A. D. Kettner et al.

(Du Pont), EP A2-0 293 881 (priority dates 5 June 1987 and 6 April 1988).

The present invention relates to novel and potent thrombin inhibitors with a competitive inhibitory action against their enzymes, i.e., they cause reversible inhibition. Another feature is the achievement of inhibitors that are orally bioavailable and selective in inhibiting thrombin, which outperforms other serine proteases. Stability, duration of action, and low toxicity at therapeutic doses are still further features of the invention.

SUMMARY OF THE INVENTION

Compounds

The compounds of the invention relate to the peptide sequence of the human fibrinogen Aa chain, representing the modified sites P _g , P ₂ and _{χ χ} ^P 9

H-Ala-Asp-Ser-Gly-Glu-Gly-Asp-Phe-Leu-Ala-Glu-Gly-Gly-Gly ^P 2 ^P 1 Ψ ^P 1 ' ^P 2' ^P 3 '

Val-Arg - Gly-Pro-Arg-ValPři.tomto invention it was found that compounds of the formula ¹ I, either as such or in the form of physiologically acceptable salts, including stereoisomers, are potent inhibitors of thrombin.

The compounds of the invention have the general formula I

in which

A is methylene or

A represents an ethylene group and the resulting five-membered ring may or may not contain one or two fluorine, hydroxy or oxo at the 4-position, or may or may not be saturated, or

A is -CH 2 -O-, -CH ₂ -S- or

-CH ₂ -SO- with heteroatom functionality at the 4-position, or

A represents an n-propylene group and the resulting six-membered ring may or may not contain one fluorine, hydroxy or oxo group at the 5-position, two fluorine atoms at one of the 4 or 5 positions, or unsaturated at the 4 and 5 positions or (C 1 -C 4); or

A represents a group of formula -CH ₂ -O-CH ₂ -, -CH ₂ -S-CH ₂ or ^- CH 2 ^- S0 ^- CH2-,

R ¹ represents a hydrogen atom, a C 1 -C 4 alkyl group, a C 2 or C 3 hydroxyalkyl group or a group of the formula R ^1: OOC-alkyl, wherein the alkyl group contains 1 to 4 carbon atoms and represents a hydrogen atom; a (C 1 -C 4) alkyl group or a (C 2 -C 3) alkylene group bonded intramolecularly to the carbonyl group at R 6, or

R ¹ is R ¹² OOC-1,4-phenyl-CH ₂ ", wherein R ¹² is hydrogen or C 1 -C 4 alkyl, or

R ² represents a group of the formula R ¹² -NH-CO-alkyl-, wherein alkyl as the alkyl group contains 1 to 4 carbon atoms and is optionally substituted at the position and to the carbonyl group by an alkyl group with 1 to 4 carbon atoms and wherein R ¹² represents a hydrogen atom or a (C 1 -C 4) alkyl group, or a group of the formula -CH ² COOR ¹² wherein R · ¹ · ² is as defined above, or

R ⁴ represents a group of the formula R 1 - OOC-C 1 - OOC-alkyl-, wherein the alkyl as alkyl group contains 1 to 4 carbon atoms and is optionally substituted at the position and to the carbonyl group with a C 1 -C 4 alkyl group and ¹² is as defined above, or

R ¹ represents a group of the formula CH 2 SO 2 -, or

R ² represents a group of formula R ¹² OCOCO-, wherein R ¹² is as defined above, or

R ¹ represents a group represented by -CH ₂ PO (OR ¹⁴ ) 2, -CH ₂ SO ₂ H or -CH ₂ - (5 (1 H) -tetrazolyl), wherein R ¹⁴ is each independently hydrogen, methyl or ethyl,

R ² represents a hydrogen atom, a C 1 -C 4 alkyl group or a group of the formula R ²¹ OOC-alkyl-, wherein the alkyl group contains 1 to 4 carbon atoms and is optionally substituted in the α-position to the carbonyl group and a substituent in the position a is a group of formula R ²² - (CH ₂ ) p - wherein p is 0 to 2 and R ²² is methyl, phenyl, hydroxy or j of COOR ²¹ and wherein R ²¹ is hydrogen or C 1-4 alkyl carbon atoms, m is 0, 1 or 2,

R ³ represents a cyclohexyl group and

R ⁴ represents a hydrogen atom, or m is 1,

R ³ represents a cyclohexyl or phenyl group and

R ⁴ forms an ethylene bridge together with R ¹ , or m represents the number 1 a

R ³ and R ⁴ are each cyclohexyl or phenyl,

R ⁵ represents a hydrogen atom or a C 1 -C 4 alkyl group, J 1 represents an integer from 2 to 6, and

B is -N (R ⁶ ) -C (NH) -NH ₂ wherein R ⁶ is hydrogen or methyl, or

Λ

B is -SC (NH) -NH ₂ or -C (NH) -NH ₂ · j

The alkyl group may have a straight or branched chain unless otherwise indicated. C 1 -C 4 alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. When unsaturation is mentioned, it is meant a carbon-carbon double bond.

The abbreviations used in some cases to denote compounds are summarized at the end of this description.

In a preferred embodiment, the present invention relates to compounds of formula I wherein R ¹ is R 10 OOC-alkyl-, wherein the alkyl group contains from 1 to 4 carbon atoms and R ¹¹ is hydrogen. Of these, compounds wherein A is ethylene and R ³ is hydrogen or C 1 -C 4 alkyl are preferred, especially those wherein R ⁵ is hydrogen.

Among the compounds of formula (I), another preferred sub-group of compound is where R ³ is cyclohexyl and m is 1 or 2, especially m is 1 and R ⁴ is hydrogen.

Another preferred group of compounds are those wherein A is n-propylene and the resulting six-membered ring may or may not have a C 1 -C 4 alkyl group at the 4-position and R ⁵ is hydrogen or C 1 -C 4 alkyl, especially those compounds wherein R ^J is hydrogen.

In another preferred embodiment, n is 3.

Compounds of formula I having the S-configuration at the α-amino acid at the P ₂ position are preferred. Of these, compounds which also have the R-configuration at the α-amino acid at the P ₃ position are particularly preferred.

Preferred compounds of the invention are:

 y

Example No. Compound

21

H - (R) Cha-Pro-Agm

Me (R) Cha-Pro-Agm

HO- (CH ₂ ) ₃ - (R) Cha-Pro-Agm

_HOOC-CH2 - (R) Cha-Pro-Agm ⁱ _PrOOC-CH2 - (R) Cha-Pro-Agm

HOOC-CH ₂ - (Me) (R) Cha-Pro-Agm

HOOC - (R, S) CH (Me) - (R) Cha-Pro-Agm

HOOC- (R or S) CH (Me) - (R) Cha-Pro-Agm / and HOOC- (R or S) CH (Me) - (R) Cha-Pro-Agm / b HOOC- (R or S) ) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm / a HOOC- (R) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm / b HOOC- (R) CH (Ph ) - (R) Cha-Pro-Agm / b HOOC- (R, S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm HOOC- (R or S) CH (CH ₂ CH ₂ Ph) ) - (R) Cha-Pro-Agm and HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Agm

EtOOC-CO - (R) Cha - Pro - Agm (R, S) - Bia - (R) Cha - Pro - Agm

HOOC- (R or S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm / b

H- (R) Cha-Pro-Nag ⁿ Bu- (R) Cha-Pro-Nag

HO- (CH ₂ ) ₃ - (R) Cha-Pro-Nag

_HOOC-CH2 - (R) Cha-Pro-Nag

EtOOC-CH ₂ - (R) Cha-Pro-Nag ⁱ _PrOOC-CH2 - (R) Cha-Pro-Nag ^t _BuOOC-CH2 - (R) Cha-Pro-Nag

HOOC-CH ₂ -OOC-CH ₂ - (R) Cha-Pro-Nag

H ₂ N-CO-CH ₂ - (R) Cha-Pro-Nag

HOOC-CH ₂ -NH-CO-CH ₂ - (R) Cha-Pro-Nag _{(HOOC-CH2) 2} - (R) Cha-Pro-Nag _HOOC-CH2 - (Me) - (R) Cha- Pro-Nag HOOC-CH ₂ - ⁽ⁿ Bu) - (R) Cha-Pro-Nag HOOC- (R, S) CH (Me) - (R) Cha-Pro-Nag HOOC- (R) CH ( Me) - (R) Cha-Pro-Nag / a .47

6

HOOC - (R or S) CH (Me) - (R) Cha - Pro - Nag / b EtOOC - (R, S) CH (Me) - (R) Cha - Pro - Nag HOOC - (R or S) CH ⁽ⁿ Pr) - (R) Cha-Pro-Nag / a HOOC- (R) CH (CH ₂ OH) - (R) Cha-Pro-Nag HOOC- (R, S) CH (Ph) - (R ) Cha-Pro-Nag HOOC- (S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Nag HOOC- (R) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Nag _{HOOC-CH2 -CH2} - (R) Cha-Pro-Nag _{EtOOC-CH2 -CH2} - (R) Cha-Pro-Nag HOOC- (CH _{2) 3} - (R) Cha-Pro-Nag EtOOC- (CH ₂ ) ₃ - (R) Cha-Pro-Nag HOOC-CO- (R) Cha-Pro-Nag

MeOOC-CO - (R) Cha - Pro - Nag (R, S) Bia - (R) Cha - Pro - Nag

HOOC- (R, S) CH (CH ₂ COOH) - (R) Cha-Pro-Nag MeOOC- (R, S) CH _(CH2 COOMe) - (R) Cha-Pro-Nag HOOC-Ph-4- CH ₂ - (R) Cha-Pro-Nag (HO) ₂ P (O) -CH ₂ - (R) Cha-Pro-Nag

EtO (HO) P (O) -CH ₂ - (R) Cha-Pro-Nag (EtO) ₂ P (O) -CH ₂ - (R) Cha-Pro-Nag

_HOOC-CH2 - (R) Cha-Pro-Mag

H- (R, S) Pro (3-Ph) -Pro-Agm

H- (R, S) Pro (3- (trans) Ch) -Pro-Agm HOOC-CH ₂ - (R, s) Pro (3- (trans) Ph) -Pro-Agm HOOC-CH ₂ - (R , S) Pro (3- (trans) Ph) -Pro-Nag _HOOC-CH2 - (R) Cha-Pic-Agm

HOOC-CH ₂ - (Me) - (R) Cha- (R, S) Pic-Agm HOOC- (R, S) CH (Me) - (R) Cha-Forge-Agm HOOC- (R or S) CH (Me) - (R) Cha-Pic-Agm / a HOOC- (R or S) CH (Me) - (R) Cha-Pic-Agm / b _{HOOC-CH2 -CH2} - (R) Cha-Pic -Agm H- (R) Cha-Pic-Nag

Me- (R) Cha- (R, S) -Pic-Nag

HOOC-CH ₂ - Cha-Pic-Nag ·

MeOOC-CH ₂ - (R) Cha-Pic-Nag

7;

69 ¹ PrOOC-CH 70 hooc-ch ₂ (i 71 HOOC- (R, S 72 HOOC- (Rn < 73 HOOC- (Rn < 74 hooc-ch ₂ - < 75 hooc-ch ₁ - 76 hooc-ch ₁ - 77 H- (R) Cha-. 78 hooc-ch ₁ - 79 H- (R) Cha-: 80 hooc-ch ₁ - 81 hooc-ch ₁ - 82 hooc-ch ₁ - 83 hooc-ch ₁ - 84 hooc-ch ₁ - 85 H- (R) Hoc- 86 hooc-ch ₁ - 87 hooc-ch ₁ - 88 hooc-ch ₁ - 89 hooc-ch ₁ - 90 hooc-ch ₁ - 91 H- (R) Cha- 92 hooc-ch ₁ - 93 hooc-ch ₁ -

Of these, compounds having the designation of Examples 4, 6, 9, 22, 30, 34, 59, 63, 67, 73, 80 and 82 are particularly preferred.

HOOC-CH ₂ - (Me) - (R) Cha-Pro-Nag

HOOC- (R or S) CH (Me) - (R) Cha-Pro-Nag / b

_HOOC-CH2 - (R) Cha-Pic-Nag

In the above compound tables, letters a, b, c and d refer to a substantially pure stereomer at the carbon atom, which is designated R or S. A streomer can be identified for each compound in connection with the experimental portion disclosed herein. R, S refers to a mixture of stereomers.

A further embodiment of the invention relates to a new use of a compound of the formula

as a starting compound for the synthesis of a serine protease inhibitor, and in particular for the synthesis of a thrombin inhibitor.

This compound can be used as such or with a guanidino group, either protected once on δ-nitrogen or twice protected on δ-nitrates or τ, δ-nitriles, preferably by a protecting group such as a benzyloxycarbonyl group. The protection of noragmatin derivatives is carried out by methods known in the art of guanidine compounds. This compound is named noragmatin or abbreviated as Nag. The compound has been previously reported, inter alia, as a hair bleach accelerator in GB Patent No. 1,599,324 (Henkel, priority date February 5, 1977). Structural fragment of the formula

however, it has not previously been described as a structural element of a pharmaceutically active compound. As such a structural element acts a noragmatin fragment in a serine protease inhibitor

and in particular acts as a valuable thrombin inhibitor.

Medical and pharmaceutical use

According to another embodiment, the present invention relates to the treatment of a human or animal organism in conditions that require inhibition of thrombin. The compounds of the invention are believed to be particularly useful in animals, including humans, for the treatment or prophylaxis of thrombosis and hypercoagulability in blood and tissues. Furthermore, it is expected that these compounds should be useful in situations where an undesired excess of thrombin is exhibited without signs of hypercoagulability. Venereal conditions in which the compounds find potential utility for treatment and / or prophylaxis include vein thrombosis, pulmonary embolism, arterial thrombosis such as myocardial infarction, unstable angina, stroke based on thrombosis, and peripheral arterial thrombosis. Further, it is expected to be used in the prophylaxis of atherosclerotic diseases such as coronary arterial diseases, cerebral arterial diseases and peripheral arterial diseases. Furthermore, the compounds are expected to be useful together with thrombolytics in thrombotic diseases, especially myocadial infarction. In addition, the compounds are believed to be useful in prophylaxis for reocclusion after thrombolysis, percutaneous transluminal angioplasty (PTCA), and coronary surgical bridges. Further, the compounds are expected to be useful in preventing retrombosis after minor surgical procedures. Still further, it is believed that the compounds are useful in anticoagulant treatment in connection with artificial organ and heart valve replacements. The compounds are further expected to be useful in anticoagulant treatment in hemodialysis and multiple intravascular coagulation.

Another expected use is in the flushing of blood vessels and mechanical devices used in patients in vivo, and as an anticoagulant for protecting blood, plasma and other blood products in vitro.

Pharmaceutical preparations

The compounds of formula (I) are usually administered by the oral, rectal, dermal, nasal or parenteral route in the form of pharmaceutical compositions which contain the active ingredient either as a free base or as a pharmaceutically acceptable non-toxic acid addition salt such as hydrochloride, hydrobromide, lactate, acetate , citrate, p-toluenesulfonate, trifluoroacetate and the like, in a pharmaceutically acceptable dosage form.

The dosage form may be a solid, semi-solid or liquid composition which is prepared in a manner known per se. Usually, the active ingredient will comprise from 0.1 to 99% by weight of the composition, in particular from 0.1 to 50% by weight for compositions intended for parenteral administration and from 0.2 to 75% by weight for compositions suitable for oral administration.

A suitable daily dose of the compounds of this invention for the therapeutic treatment of humans is about 0.001 to 100 mg / kg body weight for oral administration and from 0.001 to 50 mg / kg body weight for parenteral administration.

Method of production

A further feature of the invention is a process for the preparation of compounds. Compounds of formula I may be prepared by coupling an N-terminally protected amino acid or dipeptide or a preformed N-terminally alkylated protected dipeptide to a compound of the formula

H ₂ N- ( ^c H ₂ ) n- ^x - χο wherein ή is as defined in formula I, and

X is an unprotected or protected guanidino, protected amino or convertible to an amino group, which is subsequently converted to guanidino.

The coupling is therefore carried out in one of the following ways.

Method I

Coupling of the N-terminally protected dipeptide, produced by standard peptide coupling, with either protected or unprotected guanidine or straight chain alkylamine is performed to protect or mask the amino group at the end of the alkyl chain, using the standard peptide coupling shown in the formula

R ⁵

6 *

0 ‘

OH

H ₂ N- (CH 2) _n -X

in which samples

R ³ , R ⁴ , R ⁵ , n, m and A have the meanings given in formula I,

R ⁶ represents a hydrogen atom or an alkyl group,

W ₁ represents an amino protecting group such as tert-butoxycarbonyl and benzyloxycarbonyl; and

X is -NH-C (NH) NH ₂ , -NH-C (NH) NH-W ₂ ,

-N (W ₂ ) -C (NH) NH-W ₂ , -NH-C (NW ₂ ) NH-W ₂ or -NH-W ₂ , wherein W ₂ is an amino protecting group such as tert-butoxycarbonyl and benzyloxycarbonyl, or

X represents a masked amino group, such as an azide residue, to form a protected peptide. The final compounds can be prepared by any of the following methods, depending on the nature of the X- group used. Removal of the protecting group or groups (where X is a group of formula

-NH-C (NH) NH ₂ , -N (W ₂ ) -C (NH) NH-W ₂ , -NH-C (NW ₂ ) NH-W _2, or

-NH-C (NH) NH-W ₂ ), or selectively removing the group W ₁ (for example, when X is -NH-C (NH) NH-W ₂ , -N (W ₂ ) -C (NH) NH -W ₂ , -NH-C (NW ₂ ) NH-W ₂ , with W ₂ in this

X / case must be placed orthogonal to W 1) followed by alkylation of the N-terminal nitrogen atom and removal or selective removal or unmasking of the terminal alkylamine function (X stands for NH-W ₂ , where W ₂ in this case must be orthogonal to or X (a masked amino group such as an azide moiety) followed by a guanidization reaction is carried out using standard methods, from the free amine and removing the group.

Method II

Coupling of the N-terminally protected amino acid, produced by standard methods, with either protected or unprotected aminoguanidine or straight chain alkylamine is performed to protect or mask the amino group at the end of the alkyl chain, using the standard peptide coupling shown in the formula

R ⁵

H ₂ N- (CH 2) _n -X

- 1 «R ⁵

in which samples

C. .

A, R and X have the meanings given above, followed by removal of the group and coupling with the N-terminal amino acid, in protected form, resulting in the protected peptides described in method I or III, depending on the choice of substitution pattern on the N-atom. terminal amino acids used for coupling. The synthesis then proceeds according to Method I or Method III to produce the final peptides.

Method III

Coupling of the preformed N-terminal alkylated or protected dipeptide, produced by standard peptide coupling, with either protected or unprotected aminoguanidine or straight chain alkylamine is carried out in protecting or masking the amino group at the end of the alkyl chain, using the standard peptide coupling shown

R ³ <

W ₃

R ²

R ⁵

H ₂ N- (CH ₂ ) _n -X

NH- (CH ₂ ) _n -X in which formulas

R ^2, R ^3, R ^4, n, m, A and X are as defined above with the proviso that R ² is other than hydrogen, and a is an acyl protecting group such as trifluoroacyl.

The final compounds can be prepared by any of the following methods, depending on the nature of the group Xa used, by removing the protecting groups (wherein X- is -NH-C (NH) NH ₂ , -NH-C (NH) NH-W ₂ , -N (W ₂ ) -C (NH) NH-W ₂ ,

-NH-C (NW ₂ ) NH-W ₂ or -NH-W ₂ ) or by selective deprotection or unmasking of the terminal alkylamine function (X is -NH-W ₂ , where W ₂ in this case must be placed orthogonally k or X is a masked amino group (such as an azide moiety) followed by guanidization removal of the group W 6.

The following is a detailed description of the present invention.

The following description illustrates various aspects of the invention.

Experimental part

The synthesis of the compounds of this invention is illustrated with Schemes I to VI appended to this patent.

General experimental procedures

The @ 1 H NMR spectrum and @ ¹³ C NMR spectra are measured on a BRUKER AC-P 300 and BRUKER AM 500 spectrometer, with the BRUKER AC-P 300 spectrometer operating at a @ 1 H frequency of 500.14 MHz and a frequency of 125.76 MHz; spectrometer BRUKER AM

500 is operated at a p H frequency of 300.13 MHz and a C frequency of 75.46 MHz.

Samples weighing 10 to 50 mg are dissolved in 0.6 ml of any of the following solvents: CDCl3 (isotopic purity greater than 99.8%, Dr. Glaser AG., Basel, Switzerland), CD4 OD (isotopic purity greater than 99.8%). 99.95%, Dr. Glaser AG., Basel, Switzerland) or D ₂ O (isotopic purity higher than 99.98%, Dr. Glaser AG., Basel, Switzerland).

The values of ¹³ C chemical shifts in CDCl ₃ and CD 4 OD are relative to tetramethylsilane as an external standard.

^{The 1} H chemical shifts in D ₂ O are relative to the sodium salt of the acid. Ί Ί

The 3- (trimethylsilyl) -d ₄ -propane and C chemical shifts in D ₂ O are relative to 1,4-dioxane (67.3 ppm), both of which are external standards. Calibration with external standards may in some cases cause different small shifts compared to the internal standard, but differences in HH chemical shifts are less than 0.02 ppm

C chemical shifts of less than 0.1 ppm.

The 1 H NMR spectrum of peptide sequences containing a proline residue often shows two sets of resonances. This corresponds to the existence of contributing conformers with respect to rotation around the amide bond, wherein proline is the N-part of the nitrogen bond. The conformers are referred to as cis and trans. In the compounds of the invention, the (R) Cha-Proa (R) Cha-Pic- sequences often occur above the cis-trans equilibrium state with one conformer predominant (more than 90%). In these cases, only chemical shifts of the major rotamer are reported.

Thin layer chromatography is performed on coated glass or aluminum plates, commercially available from Merck as Silicagel 60F ₂₅₄ . Visualization is achieved by combining the effects of ultraviolet light, then spraying with a reagent solution that was made by mixing 372 mL of 95% ethanol, 13.8 mL of concentrated sulfuric acid,

4.2 ml of concentrated acetic acid and 10.2 ml of p-methoxybenzaldehyde or phosphomolybdic acid (5 to 10% by weight in 95% ethanol) and then heating.

Flash chromatography was carried out on silica gel (Silicagel 60 Merck, particle size 40-63 mm) under nitrogen overpressure.

Reverse phase high performance liquid chromatography (sometimes also referred to as RPLC) is performed on a Waters M-590 equipped with three 10Q, C8 (Eka-Nobel) reversed phase columns having different dimensions for analytical chromatography (4.6 x 250 mm), semi-preparative chromatography (2.54 cm x 250 mm) and preparative chromatography (5.08 cm x 500 mm) at 226 nm.

Freeze-drying or lyophilization is performed on a Leybold-Heraeus model Lyovac GT 2.

DETAILED DESCRIPTION OF THE INVENTION

Method of protecting compounds

Process for preparing Boc- (R) Cha-OH

To a solution of 21.55 g (125.8 mmol) of H- (R) Cha-OH in 130 ml of 1M sodium hydroxide solution and 65 ml of tetrahydrofuran was added 30 g (137.5 mmol) of (Boc) ₂₀ and the mixture was stirred at room temperature for 4.5 hours. The tetrahydrofuran was evaporated and an additional 150 ml of water was added. The alkaline aqueous phase was washed twice with ethyl acetate, then acidified with a 2 molar solution of potassium bicarbonate and extracted three times with 150 ml of ethyl acetate each. The combined organic phases were washed with water, brine and dried over sodium sulfate. Evaporation of the solvent afforded 30.9 g of the title compound as a white solid. Yield: 90.5%.

Production method of Z- (R) Cha-OH

The procedure is the same as described by M. Bodanszky and A. Bodariszky in The Practice of Peptide Synthesis, 12, Springer-Verlag (1984), starting from H- (R) Cha-OH.

Process for preparing Boc- (Me) -Phe-OH

Prepared in the same manner as Boc- (R) Cha-OH from Me- (R) Phe-OH.

Method of production. Boc- (R, S) Pro (3- (trans) Ph) -OH

To a well stirred solution of 2.0 g (8.8 mmol, 1 equivalent) of H- (R, S) Pro (3- (trans) Ph-OH x HCl (produced as described in J. Org. Chem. 55, 270-275 (1990) and J. Org Chem.

39, 1710-1716 (1974)) in 17.6 ml of 1N sodium hydroxide solution, 12 ml of water and 12 ml of tetrahydrofuran at +5 ° C are added 2.33 g (10.7 mmol, 1.2 equivalents) of (Boc) ₂ 0. the reaction mixture was allowed to warm to room temperature and stirring was continued for additional 18 hours. The organic solution was evaporated and water (50 ml) was added. The basic aqueous phase was washed twice with 50 ml of ethyl acetate each time and acidified with a 2 molar solution of potassium bicarbonate (pH about 1). The acidic aqueous phase is extracted four times with 75 ml of ethyl acetate each time and the combined organic phases are washed once with 40 ml of water, once with 40 ml of brine and dried over magnesium sulfate. Evaporation of the solvent gave 2.0 g of pure compound as a white solid. Yield: 78%.

1 H-NMR spectrum (500 MHz, CDCl ₃ , mixture of 2 rotamers): δ 1.4 and 1.5 (2s, 9H), 2.0 to 2.1 (m, 1H), 2.3 to 2 4 (m, 1H), 3.45-3.88 (m, 3H), 4.3 and 4.45 (2d, 1H), 7.2-7.4 (m,

5H) ppm.

Process for preparing Boc- (R, S) -Pro (3-Ph) -OH

Prepared as described above starting from cis / trans H- (R, S) Pro (3-Ph) -OH.

Process for preparing Boc- (R) Dph-OH

They are prepared as described by K. Hsich et al. in J. Med. Chem. 32, 898 (1989) from H- (R) Dph-OH.

Production method of Boc- (R) Hop-OH

Manufactures

Boc- (R) Cha-OH, in the same manner starting from H- (R) Hop-OH.

¹ H-NMR spectrum (300 MHz, CDCl ₃ ): δ (m, 1H), 2.22 (m, 1H), 2.75 (broad t, 2H) 1H, 5.05 (broad s, 1H) 7.15 to 7.33 (m,

1.45 (s, 9H); 2.00

4.36 (wide s,

5H) ppm.

A method of removing protecting groups

a) The protected peptide is dissolved in 95% ethanol and hydrogenated in the presence of 5% palladium on carbon at atmospheric pressure in the presence of excess trifluoroacetic acid or acetic acid greater than 2 equivalents for about 1 to 4 hours. The catalyst is filtered off, the solvent is evaporated and the final peptide (in the form of the trifluoroacetic acid or acetic acid salt) is isolated as a white powder, after removal of the water by lyophilization.

b) The procedure is the same as in a) except that the solvent is a mixture of ethanol and water in a ratio of about 5: 1.

c) Proceed as in a), except that methanol is used as the solvent.

d) Proceed as in a) except that 2-molic hydrochloric acid is used as the acid and the hydrochloride is obtained.

e) Hydrolysis of esters, exemplified as:

0.4 mmol EtOOC-CH ₂ - (R) Cha-Pro-Nag x 2 HOAc was dissolved in 1.5 mL methanol and 1.2 mL (1.2 mmol) 1 M sodium hydroxide at room temperature. After 3 hours, the methanol was evaporated and excess acetic acid was added. The mixture was lyophilized and purified by reverse phase high performance liquid chromatography using 70:30 acetonitrile: 0.1 molar ammonium acetate as eluent. The pure material is obtained as a powder in a yield of 73% of theory after lyophilization from water.

f) Cleavage of tert-butyl esters, exemplified as:

tert-butyl ester is dissolved in excess trifluoroacetic acid. The solution was stirred for 2 hours at room temperature and then the trifluoroacetic acid was evaporated. Treatment with activated carbon in aqueous ethanol is then carried out followed by lyophilization from water. This gives the desired compounds.

Production of starting compounds

Production method of H-Pic-OEt x HCl

L-pipecolinic acid (4.0 g, 0.031 mol) was suspended in absolute ethanol (100 ml) and hydrogen chloride gas was bubbled carefully through the suspension until a clear solution was obtained. The solution was cooled in an ice bath and 17 ml of thionyl chloride was added dropwise over 15 minutes. The ice bath was removed and the mixture was refluxed for 2.5 hours. The solvent was evaporated and the title compound was obtained as the hydrochloride in quantitative yield.

¹ H-NMR spectrum (300 MHz, D ₂ O): δ 1.33 (t, 3H), 1.8-2.1 (m, 5H), 2.3-2.5 (m, 1H), 3.1-3.3 (m, 1H), 3.5-3

3.7 (m, 1H), 4.14 (dd, 1H), 4.44 (q, 2H) ppm.

Production method of H-Pic-OMe x HCl

The compound was prepared in the same manner as described for H-Pic-OEt x HCl, replacing ethanol with methanol.

Production method of H-Aze-OEt x HCl

The compound was prepared in the same manner as described for H-Pic-OEt x HCl from H-Aze-OH.

Production method of H-Pic (4- (S) Me) -OEt x HCl

The compound was prepared in the same manner as described for H-Pic-OEt x HCl from H-Pic (4- (S) Me) -OH (available from Synthelec, Lund, Sweden).

Production Method of H- (R) Pic (4- (R) Me) -OEt x HCl

The compound was prepared in the same manner as described for H-Pic-OEt x HCl from H- (R) Pic (4- (R) Me) -OH (available from Synthelec, Lund, Sweden).

Production method of H- (R) Dph-OH

The compound was prepared by the general method of A. Evans et al. in JACS, 112, 4011 (1990).

Production method of H- (R, S) Pie (4,5-dehydro) -OEt

3.05 g (18.1 mmol) of H- (R, S) Pie (4,5-dehydro) -OH (produced according to the method of Burgstahler et al. In J. Org.

Chem. 25 (4), 489-492 (1960)) is dissolved in 75 ml of saturated ethanolic hydrogen chloride solution and the mixture is refluxed for 5 hours. The solvent was evaporated and the remaining residue was dissolved in water, basified with aqueous sodium hydroxide solution and extracted three times with ethyl acetate. Drying over sodium sulfate and careful evaporation gave 2.05 g of the title compound. Yield: 71%.

¹ H-NMR (CDCl ₃ ): δ 1.28 (t, 3H), 1.88 (broad s, NH), 2.2-2.4 (m, 2H), 3.45 (broad s, 2H), 3.57 (dd, 1H), 4.21 (q, 2H), 5.68-5.82 (m, 2H) ppm.

Process for preparing Boc- (R) Cgl-OH

Boc- (R) Pgl-OH was hydrogenated in the presence of 5% rhodium on alumina in methanol at 5 MPa. Filtration and evaporation of the solvent gave the title compound which was used without further purification.

1 H-NMR spectrum (300 MHz, CDCl ₃ ): δ 0.9-1.7 (m, 20H), 4.0-4.2 (m, 1H), 5.2 (d, 1H) ppm.

Process for preparing Boc- (R) Dch-OH

0.75 g (2.2 mmol) of Boc- (R) Dph-OH was dissolved in 25 ml of methanol and a catalytic amount of 5% rhodium on alumina was added. The mixture was hydrogenated at 50 MPa at 50 ° C for 40 hours. Filtration and evaporation gave 0.72 g of the title compound. Yield: 93%.

¹ H-NMR (CDCl ₃ ): δ 0.9 to 2.0 (m, 32H, of which 1.45 (broad s, 9H)), 4.55 (broad d) and 4.9 (broad d) ), two rotamers coupled for total IH, 5.7 to 6.1 (wide, NH) ppm.

Production Method of H- (R) Pro (5- (S) Me) -OMe

The compound was prepared according to the method of B. Gopalan et al. in J. Org. Chem., 51, 2405 (1986).

Production method of H-Mor-OH

The compound was prepared by the method of K. Nakajima et al. in Bull. Chem. Soc. Jpn., 51 (5), 1577-1578 (1978) and ibid. 60, 2963-2965 (1987).

Production method of H-Mor-OEt x HCl

The compound was prepared in the same manner as H-Pic-OEt x HCl from H-Mor-OH.

Method for producing Boc- (R) Cha-OSu equivalent of Boc- (R) Cha-OH, 1.1 equivalents

N-hydroxysuccinimide and dicyclohexylcarbodiimide or 1.1 equivalents of CME-CDI are dissolved in acetonitrile (about 2.5 mL / mmol acid) and stirred at room temperature overnight. The precipitate formed during the reaction is filtered off, the solvent is evaporated off and the product is dried under reduced pressure. (If CME-DCI was used in the reaction, the acetonitrile was evaporated, the residue was dissolved in ethyl acetate, and the organic phase was washed and dried. Evaporation of the solvent gave the title compound.) 1 H-NMR spectrum (500 MHz, CDCl ₃ , 2 rotamers) approximately 1: 1): δ 0.85 to 1.1 (m, 2H), 1.1 to 1.48 (m,

4H), 1.51-1.98 (m, 16H, of which 1.55 (broad s, 9H)), 2.82 (broad s, 4H), 4.72 (broad s, 1H, major rotamer) 4.85 (broad s, 1H, minor rotamer) ppm.

Boc- (Me) - (R) Cha-OSu (i) Boc- (Me) - (R) Cha-OH

A solution of 11.9 g (42.6 mmol) of Boc- (Me) - (R) Phe-OH in 150 ml of methanol was hydrogenated in the presence of 5% rhodium on alumina at 20 psi for 24 hours. Filtration of the catalyst and evaporation of the solvent gave the product as a white solid, which was used in the next step without further purification. Yield: 95%.

¹ H-NMR spectrum (500 MHz, CDCl ₃ , mixture of 2 rotamers in approximately 1: 1 ratio): δ 0.8 to 1.1 (m, 2H), 1.1 to 1.9 (m, 20H, z of which 1.47 and 1.45 (s, 9H)), 2.82 and 2.79 (s, total 3H), 4.88 and 4.67 (m, total 1H) ppm.

(ii) A method for producing Boc- (Me) - (R) Cha-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-OSu from Boc- (Me) - (R) Cha-OH.

Boc- (R) Cha-Pro-OSu (i) Boc- (R) Cha-Pro-OH

680 mmol of H- (S) Pro-H are dissolved in 750 ml of 0.87 molar sodium hydroxide solution. 170 mmol of Boc- (R) Cha-Pro-OSu dissolved in 375 ml of dimethylformamide are added dropwise over 20 minutes. The reaction mixture was stirred at room temperature for 20 modins. The mixture was acidified with a 2M potassium bicarbonate solution and extracted three times with ethyl acetate. The organic layers were combined and washed three times with water and once with brine. After drying over sodium sulfate and evaporation of the solvent, the syrupy oil is dissolved in diethyl ether, the solvent is evaporated and finally the product is dried under reduced pressure. In nearly quantitative yield, Boc- (R) Cha-Pro-OH is obtained as a white solid.

¹ H-NMR spectrum (500 MHz, CDCl-β minor rotamer):

%): δ 0.8 to 1.05 (m, 2H), 1.05 to 1.55 (m, 15H, of which 1.5 (broad s, 9H)), 1.55 to 1.8 ( m, 5H), 1.8-2.15 (m, 3H), 2.47 (m, 1H), 3.48 (m, 1H), 3.89 (m, 1H), 4.55 (m , 2H), 5.06 (m, 1H), minor rotamer signals: 2.27 (m, 1H), 3.58 (m, 1H), 4.33 (m, 1H), 5.0 (m, 1H) ppm.

(ii) A method for producing Boc- (R) Cha-Pro-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-OSu from Boc- (R) Cha-Pro-OH.

¹ H-NMR spectrum (500 MHz, CDCl ₃ , 2 5: 1 rotamers): δ 0.78 to 1.05 (m, 2H), 1.05 to 1.83 (m, 20H, thereof

1.43 (broad s, 9H)), 1.83 to 2.26 (m, 3H), 2.32 (m, 1H), 2.72 to 2.9 (m, 4H), 3.2 (m, 1H, minor rotamer), 3.52 (m 1H, major rotamer), 3.68 (m, 1H, minor rotamer),

3.89 (m, 1H, major rotamer), 4.31 (broad q, 1H, minor rotamer), 4.56 (broad q, 1H, major rotamer), 4.71 (broad t, 1H, major rotamer) 4.93 (broad t, 1H, minor rotamer), 5.22 (broad d, 1H, major rotamer),

5.44 (broad d, 1H, minor rotamer) ppm.

A method for producing Z- (R) Cha-Pro-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-Pro-OSu from Z- (R) Cha-OH.

Process for producing Boc- (R) Cha-Pic-OSt (i) Process for producing Boc- (R) Cha-Pic-OEt

6.3 g (0.023 mol) of Boc- (R) Cha-OH are dissolved in 150 ml of dichloromethane. The solution was cooled in an ice bath and 6.3 g (0.047 mol) of N-hydroxybenzotriazole and 11.2 g (0.0265 mol) of CME-CDI were added. The ice bath was removed after 15 minutes and the reaction mixture was stirred at room temperature for 4 hours. The solvent was evaporated and the residue dissolved in 150 ml of dimethylformamide and cooled in an ice bath. 4.1 g (0.021 mol) of H-PiCc-OEt x HCl was added to the solution, and the pH was adjusted to about 9 by addition of N-methylmorpholine. Remove the ice bath and stir the reaction for 3 days. The solvent was evaporated and the residue was dissolved in ethyl acetate and washed with dilute aqueous potassium bicarbonate solution, aqueous potassium bicarbonate solution and water. The organic layer was dried over sodium sulfate and evaporated. 7.7 g of Boc- (R) Cha-Pic-OEt are obtained, which product is used without further purification. Yield: 89%.

¹ H-NMR spectrum (500 MHz, CDCl 3, 2 rotamers in 3: 1 ratio): δ 0.7 to 1.0 (m, 2H), 1.1 to 1.9 (m, 29H, thereof

1.28 (t, 3H)), 1.45 (broad s, 9H), 2.01 (broad d, 1H, major rotamer), 2.31 (broad d, 1H), 2.88 (broad t, 1H, minor rotamer), 3.30 (short t, IH, major rotamer), 3.80 (broad d, IH, major rotamer), 4.15-4.3 (m, 2H), 4.5-4.7 (m, 2H, minor rotamer), 4.77 (broad q, IH, major rotamer), 4.90 (broad d, IH, minor rotamer),

5.28 (broad d, 1H, major rotamer), 5.33 (broad d, IH, major rotamer) ppm.

(ii) A method for producing Boc- (R) Cha-Pic-OH

5.6 g (0.014 mol) of Boc- (R) Cha-Pic-OEt are mixed with 100 ml of tetrahydrofuran, 100 ml of water and 7 g of lithium hydroxide. The mixture was stirred at room temperature overnight. The tetrahydrofuran was evaporated and the aqueous solution was acidified with aqueous potassium bicarbonate solution and extracted three times with ethyl acetate. The combined organic phase was washed with water, dried over sodium sulfate and evaporated. 4.9 g of Boc- (R) Cha-Pic-OH are obtained, which is used without further purification. The compound crystallizes from

I mixtures of diisopropyl ether and hexane.

1 H-NMR spectrum (500 MHz, CDCl ₃ , 2 3.5: 1 rotamers): δ 0.8 to 1.1 (m, 2H), 1.1 to 2.1 (m, 27H, z of which 1.43 (s, 9H, major rotamer), 1.46 (s, 9H, minor rotamer)), 2.33 (broad d, 1H), 2.80 (broad t, IH, minor rotamer), 3 , 33 (broad t, IH, major rotamer), 3.85 (broad d, IH, major rotamer), 4.57 (broad d, IH, minor rotamer), 4.68 (m, IH, minor rotamer), 4.77 (broad q, IH, major rotamer), 5.03 (broad s, IH, minor rotamer), 5.33 (broad d, IH, major rotamer), 5.56 (m, IH, major rotamer) ppm.

(iii) Boc- (R) Cha-Pic-OSu g (2.6 mmol) Boc- (R) Cha-Pic-OH was dissolved in 15 ml dimethylformamide at room temperature and then cooled to -18 ° C. . This temperature is maintained during the addition of the reactants. To the reaction mixture was added 0.60 g (5.2 mmol) of hydroxysuccinimide, and the reaction mixture was stirred for several minutes until the crystals dissolved. 0.56 g (2.7 mmol) of dicyclohexylcarbodiimide is dissolved in 10 ml of dimethylformamide and the previously cooled solution is added dropwise to the reaction mixture. After keeping the reaction mixture at -18 ° C for several minutes, the reaction mixture is placed in a water bath at 20 ° C for 2 hours with stirring. The organic phase is washed once with water, twice with 0.3 molar potassium hydrogen carbonate solution, twice with dilute potassium hydrogen carbonate solution, once with water and once with sodium chloride solution and then dried with sodium sulfate. The solvent was evaporated under reduced pressure to give 1.16 g of the title compound. Yield: 93%. According to the ¹ H-NMR spectrum, this compound contains two diastereomers (epimers in Pie, S / R) in a ratio of 95: 5.

¹ H-NMR spectrum (300 MHz, CDCl ₃ , major diastereomer): δ 0.7 to 2.0 (m, 27H, of which 1.46 (broad s, 9H)), 2.29 (broad d, 1H), 2.85 (br s, 4H), 3.40 (m,

1H), 4.5-4.8 (m, 1H), 5.1-5.4 (m, 1H), 5.70 (broad d, 1H, major rotamer) ppm.

Process for the preparation of Boc- (R) Cha-Mor-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OSu from H-Mor-OEt x HCl except that acetonitrile was used as a solvent instead of dimethylformamide in the manufacture of the OSu-ester.

Process for preparing Boc- (Me) - (R) Cha-Pro-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-Pro-OSu from Boc- (Me) - (R) Cha-OH.

Process for preparing Boc- (Me) - (R) Cha-Pic-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OSu from Boc- (Me) - (R) Cha-OH.

Process for preparing Boc- (R, S) Pro (3-Ph) -Pro-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-Pro-OSu from Boc- (R, S) Pro (3-Ph) -OH.

Process for the preparation of Boc- (R, S) Pro (3- (trans) Ph) -Pro-OSu (i) Process for the preparation of Boc- (R, S) Pro (3- (trans) Ph) -Pro-OBn

To a suspension of 1.0 g (3.43 mmol, 1 equivalent)

Boc- (R, S) Pro (3- (trans) Ph) -OH, 1.04 g (4.29 mmol, 1.25 equivalents) H-Pro-OBn x HCl and 0.04 g (0.24 mmol, 0.07 equivalents) of N-hydroxybenzotriazole in 15 mL of dimethylformamide was added 1.83 g (4.29 mmol, 1.25 equivalents) of CME-CDI and 0.525 mL (4.73 mmol, 1.38 equivalents) of N- methylmorpholine at room temperature. After stirring for an additional 4 days, the solvent was evaporated and the residue was taken up in 200 ml of ethyl acetate. The organic phase is washed twice with 40 ml of water, twice with 25 ml of a 1 molar solution of potassium hydrogen sulphate, twice with 25 ml of a 1 molar solution of sodium hydroxide, twice with 25 ml of water and dried over magnesium sulphate. Evaporation of the solvent and flash chromatography, eluting with dichloromethane / methanol (97: 3) gave the pure compound as a diastereomer mixture in a ratio of about 1: 1. Yield: 44%.

(ii) A method for producing Boc- (R, S) Pro (3- (trans) Ph) -Pro-OH

The benzyl ester from the compound of the previous step was removed by hydrogenation using 5% palladium on carbon in ethanol at atmospheric pressure for 4 hours. Filtration and evaporation gave the pure material as a 1: 1 mixture of diastereomers in quantitative yield.

¹ H-NMR spectrum (500 MHz, CDCl 3, 2 diastereomers, each consisting of 2 rotamers): δ 1.3 to 2.4 (m + 4s from Boc groups, total 14H), 2.5 to 2.9 ( m, total 1H), 3.2 to 3.9 (m, total 5H), 4.3 to 4.65 (m, total 2H), 7.2 to 7.5 (m, 5H) ppm.

(iii) Method for producing Boc- (R, S) Pro (3- (trans) Ph) -Pro-OSu. The compound is prepared according to the method described for Boc- (R) Cha-OSu from Boc- (R, S) Pro. (3- (trans) Ph) -Pro-OH.

Process for the preparation of Boc- (R, S) Pro (3- (trans) Ch) -Pro-OSu (i) Process for the preparation of Boc- (R, S) Pro (3- (trans) Ch) -Pro-OH

Boc- (R, S) Pro (3- (trans) Ph) -Pro-OH is hydrogenated in the presence of 5% rhodium on alumina in methanol, in the presence of a small amount of acetic acid for 7 days at 50 psi. The catalyst was filtered off, the solvent was evaporated and the residue subjected to flash chromatography using a 94: 6 mixture of dichloromethane and methanol as eluent. The pure compound is obtained as a mixture of two diastereomers as a white solid.

(ii) A method for producing Boc- (R, S) Pro (3- (trans) Ch) -Pro-OSu

The compound was prepared according to the method described for Boc- (R) Cha-OSu from Boc- (R, S) Pro (3- (trans) Ph) -Pro-OH.

Boc- (R) Hoc-Pro-OH (i) Boc- (R) Hoc-OH

3.2 g (11.46 mmol) of Boc- (R) Hop-OH were dissolved in 75 mL of methanol. 0.5 g of rhodium on activated alumina was added to the solution, and the mixture was stirred under a hydrogen atmosphere at 50 psi for 18 hours. The catalyst was filtered off through a pad of celite (diatomaceous earth) and the solvent was evaporated.

This gives the desired compound in almost quantitative yield.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.90 (m, 2H),

1.08 to 1.33 (m, 6H), 1.43 (s, 9H), 1.60 to 1.74 (m, 6H),

1.88 (broad s, 1H), 4.27 (broad s, 1H) ppm.

(ii) A method for producing Boc- (R) Hoc-OSu

The compound was prepared in the same manner as described for Boc- (R) Cha-OSu from Boc- (R) Hoc-OH.

(iii) A method for producing Boc- (R) Hoc-Pro-OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pro-OH from Boc- (R) Hoc-OSu.

1 H-NMR spectrum (500 MHz, CDCl 3): δ 0.80 to 0.94 (m,

2H), 1.05 to 1.36 (m, 7H), 1.36 to 1.48 (broad s, 9H), 1.48 to 1.78 (m, 7H), 1.98 to 2.14 (m, 2H), 2.34 (m, 1H), 3.48 (m 1H), 3.85 (m, 1H), 4.43 (m, 1H), 4.52 (broad d, 1H) 5.26 (broad d, 1H), minor rotamer signals are apparent at δ 1.92, 2.25, 3.58, 4.20 and 4.93 ppm.

Boc- (R) Hoc-Pic-OH (i) Boc- (R) Hoc-Pic-OMe

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OEt from Boc- (R) Hoc-OH and H-Pic-OMe x HCl.

(ii) A method for producing Boc- (R) Hoc-Pic-OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from Boc- (R) Hoc-Pic-OMe.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ); δ 0,82 to 0,97 (m,

2H), 1.10 to 1.36 (m, 7H), 1.36 to 1.50 (broad s, 9H), 1.50 to 1.82 (m, 11H), 2.35 (broad d, 1H), 3.28 (broad t, 1H), 3.85 (broad d, 1H), 4.63 (m, 1H), 5.33 (broad s, 1H), 5.44 (broad d, 1H) ), minor rotamer signals are evident at δ 1.88, 2.80, 4.25, 4.55 and 4.97 ppm.

Process for preparing Boc- (R) Cha-Aze-OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from H-Aze-OEt x HCl.

Production method of Boc- (R) Cha-Pic (4- (S) Me) -OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from H-Pic (4- (S) Me) -OEt x HCl except that dichloromethane was used as the solvent.

Process for the preparation of Boc- (R) Cha- (R) Pic (4- (R) Me) -Su (i) Process for the preparation of Boc- (R) Cha- (R) Pic (4- (R) Me) -OEt

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OEt from H- (R) Pie (4- (R) Me) -OEt x HCl.

(ii) A method for producing Boc- (R) Cha- (R) Pie (4- (R) Me) -OH

The compound is prepared by applying the deprotection method (e) to the product (i) above.

(iii) A method for producing Boc- (R) Cha- (R) Pie (4- (R) Me) -OS

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OSu from Boc- (R) Cha- (R) Pic (4- (R) Me) -OH.

Process for preparing Boc- (R) Cha- (R, S) Pic (4,5-dehydro) - OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from H- (R, S) Pie (4,5-dehydro) -OEt.

Boc- (R) Cgl-Pic-OH (i) Boc- (R) Cgl-Pic-OMe

1000 ml (8.1 mmol) of pivaloyl chloride was added to a solution of 2.086 g (8.1 mmol) of Boc- (R) Cgl-OH and 1.13 ml (8.1 mmol) of triethylamine in a mixture of 25 ml of toluene and 5 ml of dimethylformamide. and then 1.46 g (8.1 mmol) of H-Pic-OMe x HCl and 1.13 ml (8.1 mmol) of triethylamine in 20 ml of dimethylformamide on an ice bath are added successively. The reaction mixture was allowed to warm slowly to room temperature and after 24 hours it was diluted with water and extracted with toluene. After washing with 0.3M KHSO4, 10% Na2CO3 and brine, the solvent was evaporated under reduced pressure to give 2.52 g of a colorless oil which was used without further purification. Yield: 81%.

1 H-NMR spectrum (500 MHz, CDCl ₃ , 2 rotamers in 5: 1 ratio): δ 0.8 to 1.8 (m, 25H), 2.25 (d, 1H), 2.75 (t, 1H, minor rotamer), 3.3 (t, 1H), 3.7 (s, 3H), 3.85 (d, 1H),

4.3 (t, 1H, minor rotamer), 4.5 to 4.6 (m, 1H), 5.25 (d, 1H), 5.30 (d, 1H) ppm.

(ii) A method for producing Boc- (R) Cgl-Pic-OH

The compound is prepared according to the method described for the hydrolysis of Boc- (R) CHa-Pic-OEt using the compound of part (i) above. The material was crystallized from a mixture of diisopropyl ether and hexane.

1 H-NMR spectrum (500 MHz, CDCl ₃ , 2 rotamers in 5: 1 ratio): δ 0.8 to 1.8 (m, 25H), 2.3 (d, 1H), 2.8 (t, IH minor rotamer), 3.3 (t, 1H), 3.9 (d, 1H), 4.4 (t, 1H, minor rotamer), 4.5 to 4.6 (m, 1H), 5, 1 (s, 1H, minor rotamer), 5.3 (d, 1H), 5.40 (d, 1H) ppm.

Process for the preparation of Boc- (R) Dph-Pic-OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from Boc- (R) Dph-OH.

Process for the preparation of Boc- (R) Dch-Pic-OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from Boc- (R) Dch-OH.

Production method of Boc- (R) Cha-Pro (5- (S) Me) -OH

The compound was prepared in the same manner as described for Boc- (R) Cha-Pic-OH from H-Pro (5- (S) Me) -OMe.

Process for the preparation of Boc-Nag (Z) (i) Process for the preparation of N-benzyloxycarbonyl-O-methylurea

To a stirred solution of 2.8 liters (53 mol) of 50% by weight aqueous sodium hydroxide solution and 32 liters of water at 18 ° C was charged in two portions a total of 1.7 kg (13.0 mol) of 94% O- methylisourea and 1.57 kg (9.0 mol) of 99% O-methylisourea hydrogen sulphate. The reaction mixture is cooled to 3-5 ° C and 3.88 kg (20.9 mol) of 92% benzyl choroformate is added over 20 minutes with cooling and vigorous stirring. During the addition of Z-Cl, the temperature of the reaction mixture was adjusted from 3 to 8 ° C. The dropping funnel is rinsed with 5 liters of water which is added to the reactor. The reaction mixture was stirred at 0-3 ° C for 18 hours, filtered and the crystals were washed with 10 L of cold water at 3 ° C. The reaction mixture is dried under reduced pressure of 1 to 2 kPa at 25 ° C for 48 hours. This gives 3.87 kg of the title compound as a white crystalline powder. Yield: 89%.

(ii) Method of Manufacturing Boc-Nag (Z)

To a stirred solution of 3.9 kg (18.5 mol)

Boc-NH- (CH ₂ ) 3 ^- NH ₂ x HCl (produced as reported by PG Mattingly in Synthesis, 367 (1990)) in 24 kg isopropane at 60-70 ° C is added portionwise over 30 minutes 4.2 kg (42 mol) potassium bicarbonate. A slow release of carbon dioxide gas occurs. The mixture was stirred for an additional 30 minutes and then N-benzyloxycarbonyl-O-methylurea (3.741 kg, 18.0 mol) was added portionwise over 30 minutes. The reaction mixture was stirred at 65-70 ° C for 16 hours, cooled to 20 ° C, and filtered. The precipitate was washed with 10 and then 5 liters of isopropanol. The combined filtrates are evaporated under reduced pressure while maintaining the heating mantle at a temperature not higher than 65 to 70 ° C. When approximately 45 liters of liquid were distilled off, 90 liters of ethyl acetate were added. The reaction mixture is cooled to 20-25 ° C, washed with 10 and 5 liters of water and 5 liters of sodium chloride solution and dried with 2 kg of sodium sulfate.

The reaction mixture was stirred, then filtered, and the filter cake was washed with 11 and 7 liters of ethyl acetate. The combined filtrates are evaporated under reduced pressure while maintaining the heating mantle at a temperature not higher than 40 to 50 ° C. When about 90 liters of ethyl acetate were distilled off, 25 liters of toluene were added and the evaporation was continued. After approximately 18 liters of distillate were collected, 20 liters of toluene were added with vigorous stirring, and the resulting mixture was cooled to -1 to 0 ° C and stirred gently overnight for 17 hours. The crystalline suspension was filtered and the obtained substance was washed with 10 and 5 liters of cold toluene. Drying under reduced pressure, at 1 to 2 kPA, at 40 ° C for 24 hours, yields 4.83 kg (13.8 mol) of Boc-Nag (Z). Yield: 76%.

¹ H-NMR spectrum (300 MHz, CDCl ₃ ): δ 1.41 (s, 9H), 1.6-1.7 (m, 2H), 3.0-3.3 (m, 4H), 4 , 8 to 5.0 (wide s,

1H), 5.10 (s, 2H), 7.2-7.4 (m, 5H) ppm.

Production method of Boc-Agm (Z) (i) Production method of Boc-Agm

To a suspension of 14.95 g (65.5 mmol, 1 equivalent) of agmatine sulfate (Aldrich), 13.7 mL (98.25 mmol, 1.5 equivalents) of triethylamine, 165 mL of water and 165 mL of tetrahydrofuran was added 21.5 g. (98.25 mmol, 1.5 equivalents) of (Boc) ₂₀ at room temperature over 5 minutes. The mixture was stirred vigorously overnight, evaporated to dryness and washed twice with 100 ml of diethyl ether. Boc-Agm is obtained as a white powder which is used without further purification in the next step.

(ii) Method of Production of Boc-Agm (Z)

To a cold suspension (+ 5 ° C) about 65.5 mmol of crude Boc-Agm from the previous step in 180 ml

Of 4N sodium hydroxide solution and 165 ml of tetrahydrofuran was added 24 ml (169 mmol, 2.5 equivalents) of benzyl chloroformate over 10 minutes. The reaction mixture was stirred at room temperature for 4 hours, then 150 mL was added. of methanol and stirring at room temperature was continued for another 20 hours. The organic solvent was evaporated and water (200 ml) was added. The basic aqueous phase is extracted once with 300 ml and then twice with 200 ml of ethyl acetate. The combined organic phases were washed twice with 100 ml of water and once with 100 ml of brine and dried over magnesium sulfate. Evaporation of the solvent and flash chromatography using a 97: 3, 95: 5 and 9: 1 gradient of dichloromethane / methanol as eluent gave 14.63 g of pure Boc-Agm (Z) as a white powder. Yield 58%.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 1.35-1.40 (m,

2H), 1.45 (s, 9H), 1.5-1.6 (m-2H), 3.0-3.2 (m, 4H),

4.65 (broad s, 1H), 5.1 (s, 2H), 7.25 to 7.40 (m, 5H) ppm.

¹³ C-NMR (75.5 MHz, CDCl ₃ ): δ 25.44, 27.36, 28.21, 65.83, 79.15, 127.47, 127.66, 128.14, 137, 29, 156.47, 161.48, 163.30 ppm.

Process for the preparation of Boc-NH- (CH ₂ ) ₃ -N ₃

The compound was prepared as described by P. G. Mattingly in Synthesis, 367 (1990).

Production method Ζ-ΝΉ- (CH ₂ ) ₂ -NH ₂

To a cold solution of 6 g (0.1 mol) of ethylenediamine and 22 ml of triethylamine in 20 ml of chloroform was added 2.5 g of Z-OSu dissolved in 5 ml of chloroform. The mixture was allowed to warm to room temperature and then allowed to stir overnight. Filtration, evaporation of the solvent, and flash chromatography, eluting with 95: 5 dichloromethane / methanol saturated with ammonia gave 0.9 g of the title compound. Yield: 46%.

1 H-NMR spectrum (300 MHz, CDCl ₃ ): δ 1.27 (s, 2H),

2.85 (t, 2H), 3.24 (q, 2H), 5.14 (s, 2H), 7.22-7.40 (m,

5H) ppm.

Production method Agm x HCl

The compound is made from Agm x H ₂ SC> ₄ (Aldrich) by exchange of> hydrogen sulfate ion with chloride ion on an ion exchange column.

Production method of H-Nag (Z) x 2 HCl

The compound is prepared by bubbling hydrogen chloride (e) with a solution of Boc-NAg (Z) in ethyl acetate and then evaporating the solvent.

BnOOC-CH ₂ -NH-CO-CH ₂ -Br

To a solution of 5 mmol of H-Gly-OBn x p-TsOH and 5 mmol of triethylamine in 10 mL of dichloromethane was added 5 mmol of 2-bromoacetic acid dissolved in 10 mL of dichloromethane and 5 mmol of dicyclohexylcarbodiimide. The mixture was stirred at room temperature overnight and filtered. The organic phase is washed twice with a 0.2 molar potassium bicarbonate solution, a 0.2 molar sodium hydroxide solution, a sodium chloride solution and dried. Evaporation and flash chromatography, eluting with a 95: dichloromethane / methanol mixture:

5, the desired compound is obtained in quantitative yield.

¹ H-NMR spectrum (300 MHz, CDCl ₃ ): δ 3.89 (s, 2H),

4.05-4.11 (d, 2H), 5.19 (s, 2H), 7.06 (broad s, 1H), 7.3-7.4 (m, 5H) ppm.

BnOOC-CH ₂ -OCO-CH ₂ -Br

A mixture of 2.8 g (0.020 mmol) of bromoacetic acid, 4.2 g (0.020 mmol) of benzyl bromoacetate and 2.0 g (0.020 mmol) of triethylamine in 25 ml of ethyl acetate was refluxed for 3 hours. The reaction mixture was then diluted with more ethyl acetate and cooled. The solution was washed with dilute hydrochloric acid and then with aqueous potassium bicarbonate solution and finally with water. Drying over sodium sulfate and evaporation followed by flash chromatography (75/25 heptane / ethyl acetate) gave the title compound (26%).

¹ H-NMR spectrum (300 MHz, CDCl ₃ ): δ 3.95 (s, 2H),

4.75 (s, 2H), 5.23 (s, -2H), 7.35 to 7.45 (m, 5H) ppm.

Production method of BnO- (CH ₂ ) ₃ -OTf

Propanediol monobenzyl ether (0.83 g, 5 mmol) was dissolved in a mixture of dry pyridine (0.6 g, 7 mmol) and dichloromethane (20 ml) and cooled to -15 ° C.

Trifluoromethanesulfonic anhydride, previously cooled to -15 ° C, was added to the reaction mixture, and the resulting mixture was stirred for 45 minutes while allowing the temperature to rise to 15 ° C. The solvent was evaporated and the residue was dissolved in 10 ml of 4: 1 hexane-ethyl acetate. The solution was filtered through silica. The solvent was finally evaporated and the residue was dried under reduced pressure. 0.95 g of 1-benzyloxy- (3-trifluoromethanesulfonylpropane) is obtained, which is used directly further (see Example 21). Yield: 64%.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 2.12 (m, 2H), 3.6 (t, 2H), 4.51 (s, 2H), 4.72 (t, 2H), 7.22 to 7.42 (m, 5H) ppm.

Process for preparing BnO- (CH ₂ ) ₂ -CHO

The compound is prepared by Swern oxidation as described by D. Swern et al. in J. Org. Chem. 2480 - 2482 (1978)

Of BnO- _{(CH2) 3} -OH.

¹ H-NMR spectrum (300 MHz, CDCl ₃ ): δ 2.63 (dt, 2H),

3.80 (t, 2H), 4.51 (s, 2H), 7.30 (m, 5H), 9.76 (broad t,

1H) ppm.

Method Br- (S) CH (CH ₂ 0BN) -COOBn (i) Method of Br- (S) CH (CH ₂ 0BN) -COOH

3.9 g (19 mmol) of O-benzylserine in 10 ml of water are added to a solution of 11 g (107 mmol) of sodium bromide in 20 ml of water and 2 g (20 mmol) of sulfuric acid. The reaction mixture was cooled to -10 ° C and 1.73 g (25 mmol) of sodium nitrite was added with vigorous stirring. An additional portion of water was added to the thick mixture, and after a few minutes 1 g (10 mmol) of sulfuric acid was added. The mixture was stirred at room temperature overnight and then extracted twice with 100 ml of ethyl acetate each time. The combined organic phases were washed twice with water and once with brine and dried over sodium sulfate. Evaporation of the solvent gave

3.7 g of the title compound as a yellow oil which is pure enough to be used directly in the next step. Yield: 75%.

(ii) Production method of Br- (S) CH (CH ₂ O Bn) -COOBn

To a solution of 2.6 g (10 mmol) of the crude compound from part (i) above in 25 ml dry benzene was added 2.6 g (20.5 mmol) oxalyl chloride and 1 g molecular sieve 4 (4 x 10 ^-10 m). ). The mixture was stirred at room temperature under an argon atmosphere for 18 hours. The molecular sieve was filtered off and the solution was evaporated. The light yellow residue was dissolved in 10 mL of acetonitrile and 1 g (9.2 mmol) of benzyl alcohol was added. The resulting mixture was stirred at room temperature for 5 hours. The solvent was evaporated and the residue was dissolved in diethyl ether and washed once with 1M sodium hydroxide solution, water and brine and then dried over sodium sulfate. Evaporation of the solvent followed by flash chromatography (95: 5 dichloromethane: methanol) gave 1.8 g of the title compound. Yield: 67%.

1 H-NMR spectrum (500 MHz, CDCl 3): δ 3.82 (dd, 1H),

3.99 (dd, 1H), 4.38 (dd, 1H), 4.56 (s, 2H), 5.23 (s, 2H),

7.23 to 7.46 (m, 5H) ppm.

Working examples

Example 1

Production method of H- (R) Cha-Pro-Agm x 2 HOAc (i) Production method of Boc- (R) Cha-Pro-Agm x HOAc

1.7 mmol of Boc- (R) Cha-Pro-OSu and 2.0 mmol (1.18 equivalents) of agmatine dihydrochloride are dissolved in 35 ml of a 95: 5 mixture of dimethylformamide and water. Triethylamine was added to adjust the pH to about 10, and the solution was stirred at room temperature for 2 days. The solution was evaporated to dryness at 60 ° C / 665 Pa and the crude material was purified by reverse phase high performance liquid chromatography, eluting with a 38:62 mixture of acetonitrile and 0.1 molar ammonium acetate. The title compound was obtained as a white solid. powder after lyophilization.

spectrum (500 MHz, 5: 2 mixture of CDClg and DMSO-dg, two 9: 1 rotamers): S (majority rotamer): 0.75 to 0.90 (m, 2H), 1.1 to 2 .05 (m, 19H), 1.35 (s, 9H), 2.98-3.14 (m, 4H), 3.37 (q, 1H), 3.76 (m, 1H),

4.20 (m, 1H), 4.33 (dd, 1H), 6.30 (d, 1H), 7.05 to 7.80 (broad m, 5H), 8.67 (broad d, 1H) ppm.

Changes in the minor minor rotamer signals are observed obscure at δ 3.44 (m, 1H), 3.62 (m, 1H), 4.10 (m, 1H), 4.64 (m, 1H), 5.56 ( d, 1H), 9.08 (m, 1H) ppm.

(ii) A method for producing H- (R) Cha-Pro-Agm x 2 HOAc

A solution of 0.2 mmol Boc- (R) Cha-Pro-Agm in 2 mL trifluoroacetic acid was stirred at room temperature for 4.5

I hours. The solvent was evaporated and the remaining oil was subjected to reverse phase high performance liquid chromatography, eluting with a 25:75 mixture of acetonitrile and 0.1 molar ammonium acetate. After repeated lyophilization, the diacetate salt is obtained in the form of a white powder.

1 H-NMR spectrum (500.13 MHz, D ₂ O): δ 0.80 to 0.95 (m, 2H), 1.00 to 1.21 (m, 3H), 1.32 (m, 1H) 1.40 to 1.78 (m, 12H), 1.83 to 2.00 (m, 2H), 1.90 (s, acetate), 2.20 (m, 1H), 3.06 to m 3.14 (m, 4H), 3.50 (m, 1H), 3.67 (m, 1H), 4.20-4.30 (m, 2H) ppm.

¹³ C-NMR (75.6 MHz, D ₂ O): guanidine: δ 157.4, carbonyl carbons: δ 169.9, 174.5 ppm.

Example 2

Production method of Me- (R) Cha-Pro-Agm x 2 HOAc (i) Production method of Boc- (Me) - (R) Cha-Pro-Agm

To a solution of 479.6 mg (1 mmol, 1 equivalent) of Boc- (Me) - (R) Cha-Pro-OSu and 500 ml of N-methylmorpholine in 16 ml of a 15: 1 mixture of dimethylformamide and water was added 166 0.5 mg (1.2 mmol, 1.2 equivalents) of agmatine hydrochloride at room temperature. The reaction mixture was stirred for an additional 70 hours and the solvent was evaporated. The crude compound is obtained as an oil. This material was used in the next step without further purification.

(ii) Production method of (Me) - (R) Cha-Pro-Agm x 2 HOAc

The crude oil from the preceding step was dissolved in 10 mL of trifluoroacetic acid. and dichloromethane 1: 4 at room temperature. After stirring for 2 hours 25 minutes, the solvent is evaporated and the crude material is purified by reverse phase high performance liquid chromatography, eluting with a mixture of acetonitrile and 0.1 molar ammonium acetate (35:65). compound as a white powder.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 0.93-1.05 (m,

2H), 1.10 to 1.29 (m, 3H), 1.33 to 1.43 (m, 1H), 1.50 to

1.80 (m, 12H), 1.88-2.10 (m, 2H), 1.92 (s, acetate), 2.27-2.36 (m, 1H), 2.68 (s, 3H), 3.15-3.23 (m, 3H), 3.24-3.31 (m, 1H), 3.57-3.66 (m, 1H), 3.76-3.88 (m, 1 H),

4.28 (t, 1 H), 4.39 (dd, 1 H) ppm.

¹³ C-NMR (125.76 MHz, D ₂ O): guanidine: δ 157.24, carbonyl carbons: δ 174.03, 168.24 ppm.

Example 3

Method for producing HO- (CH ₂ ) ₃ - (R) Cha-Pro-Agm x 2 HCl (i) Method for producing Boc- (R) Cha-Pro-Agm (Z) equivalent to Boc-Agm (Z) is dissolved in the mixture trifluoroacetic acid and dichloromethane 1: 4 (about 6 mL / mmol) and stirred at room temperature for about 2 hours. The solvent was evaporated and the residue was dissolved together with 1 equivalent of Boc- (R) Cha-Pro-OSu in dimethylformamide (about 1 mL / mmol), the pH was adjusted to about 9 by addition of N-methylmorpholine, and the mixture was stirred at room temperature for for 20 hours. The solvent was evaporated under reduced pressure and the crude material was dissolved in dichloromethane and washed three times with water and once with brine. After the solution was dried over sodium sulfate, the solvent was evaporated and the compound was subjected to flash chromatography, eluting with a mixture of dichloromethane and methanol. So take the form of white powder £?

gets Boc- (R) Cha-Pro-Agm (Z).

(ii) Production method of H- (R) Cha-Pro-Agm (Z)

Boc- (R) Cha-Pro-Agm (Z) was dissolved in a 1: 4 mixture of trifluoroacetic acid and dichloromethane (approximately 6 mL / mmol) and stirred at room temperature for 2 hours. The solvent was evaporated and the residue was dissolved in 0.2 M sodium hydroxide solution (20 ml / mmol) and extracted twice with dichloromethane. The organic layers were combined and washed with brine, dried over sodium sulfate, and then the solvent was evaporated. H- (R) Cha-Pro-Agm (Z) is obtained as a white powder.

(iii) Preparation of BnO- (CH ₂ ) ₃ - (R) Cha-Pro-Agm (Z) mmol of H- (R) Cha-Pro-Agm (Z) was dissolved in 10 mL of methanol. To the solution was added 1 mmol of triethylammonium hydrochloride, 0.7 mmol of sodium cyanoborohydride, and then 1.05 mmol of BnO- _(CH2) 2 CHO ^_ and the reaction mixture stirred at room temperature overnight. The solvent was evaporated and the crude material was dissolved in ethyl acetate, washed twice with water, once with brine and dried over sodium sulfate. The solvent was evaporated and the crude material was purified by flash chromatography, eluting with a mixture of ethyl acetate and methanol.

(iv) A method for producing HO- (CH ₂ ) ₃ - (R) Cha-Pro-Agm x 2 HCl

The compound is produced by applying process (d) of deprotection to the product (iii) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 0.72 (m, minor rotamer), 0.84 (m, minor rotamer),. 0.87 to 1.03 (m, 2H),

1.03 to 1.26 (m, 3H), 1.28 to 1.40 (broad s, 1H)

1.80 (m, 11H), 1.80-1.95 (wide s, 3H), 1.95-2.10 (wide s, 2H), 2.28 (m, 1H), 3.04 ( m, 1H), 3.08-3.3 (m,

5H), 3.58 (broad s, 1H), 3.67 (broad s, 2H), 3.78 (m, 1H), 4.12 (broad d, minor rotamer), 4.30 (m, IH) ), 4.37 (m,

1H) ppm.

¹³ C-NMR (125 MHz, D ₂₀ ): guanidine: δ 157.26, carbonyl carbons: δ 174.06, 168.36 ppm.

Example 4

Production method of HOOC-CH ₂ - (R) Cha-Pro-Agm x HOAc

A general method for alkylating a terminal nitrogen atom

This method is described in connection with the more general data and will be exemplified below, together with the alkylating agent used in each particular example.

equivalent of the peptide to be alkylated and 1.1 to 1.2 equivalents of the alkylating agent are dissolved in about 10 mL / mmol of acetonitrile. Potassium carbonate (2.0-2.2 equivalents) was added to the solution, and the reaction mixture was stirred at 50-60 ° C until the starting material was needed (usually by 1-5 hours according to thin layer chromatography). Filtration, evaporation of the solvent and flash chromatography, eluting with a mixture of dichloromethane and methanol, dichloromethane and methanol saturated with ammonia or ethyl acetate and methanol in a ratio of about 9: 1, gives the alkylated substance after evaporation of the solvent.

(i) BnOOC-CH ₂ - (R) Cha-Pro-Agm (Z) production method

The compound was prepared from H- (R) Cha-Pro-Agm (Z) (see Example 3) and Br-CH ₂ COOBn according to the method described above.

(ii) Production method of HOOC-CH ₂ - (R) Cha-Pro-Agm x HOAc

The compound is produced by applying process (b) of deprotection to the product (i) above.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.9-1.1 (m, 2H), 1.1-2.3 (m, 19H), 1.95 (s, acetate), 3, 1 to 3.2 (m, 4H), 3.2 to 3.65 (m, 3H), 3.85 (m, 1H), 4.0 (broad t, 1H), 4.35 (dd, IH) ) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.55, carbonyl carbons: δ 168.71, 171.37 and 174.3 ppm.

Example 5

Production Method ¹ Pr-OOC-CH ₂ - (R) Cha-Pro-Agm x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-Agm (Z) (see Example 3) and Br-CH ₂ COO ¹ Pr, followed by deprotection according to method (b). The title compound is obtained.

1 H-NMR spectrum (300 MHz, MeOD): δ 0.85-1.05 (m, 2H), 1.1-1.35 (m, 9H, thereof 1.23 (d, 3H), 1 25 (d, 3H)), 1.35-2.02 (m, 14H), 1.92 (s, acetate), 2.08 (m, 1H), 2.2 (m, 1H), 3 07-3.45 (m, 6H), 3.55 (m, 1H), 3.7-3.8 (m, 2H), 4.3 (dd, 1H), 5.05 (m, 1H) ) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.39, carbonyl carbons: δ 171.10, 172.76 and 174.44 ppm.

Example 6

Production method of HOOC-CH ₂ - (Me) - (R) Cha-Pro-Agm x 2 TFA (i) Production method of Me- (R) Cha-Pro-Agm (Z)

The compound was prepared from Boc- (Me) - (R) Cha-Pro-OSu in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3.

(ii) Production method of HOOC-CH ₂ - (Me) - (R) Cha-Pro-Agm x 2 TFA

It is alkylated as in Example 4, using Me- (R) Cha-Pro-Agm (Z) and Br-C 1 -C 4 COOBn followed by deprotection according to method (b). The title compound is obtained.

¹ H-NMR spectrum (300 MHz of D ₂ 0): δ 0,9 to 1.35 (m, 6H) , 1, 5-2.2 (m, 14H), 2.25 to 2.45 (m, 1H), 3.12 (s, 3H) 15-3.35 (m, 4H), 3.6 to 3.75 (m, 1H), 3.8 to 3.95 (m, 1H) , 4.22 (seemingly wide with, 2H), 4.45 (m, 1H), 4.6

(broad t, 1 H) ppm.

¹³ C-NMR (75.47 MHz, D ₂ 0): guanidine: δ 157.52 and carbonyl carbons: δ 173.86, 168.79, 167.38.

Example 7

Production method of HOOC- (R, S) CH (Me) - (R) Cha-Pro-Agm. x HOAc

It is alkylated as in Example 4, using H- (R) Cha-Pro-Agm (Z) (see Example 3) and Br-CH (Me) COOBn followed by deprotection according to method (a). The title compound is obtained as a mixture of 2 diastereomers.

Example 8

Production method of HOOC- (R or S) CH (Me) - (R) Cha-Pro-Agm / a x HOAc

The compound is obtained by resolution of the diastereomers formed in Example 7, using high performance reverse phase liquid chromatography, eluting with a 1: 4 mixture of acetonitrile and 0.1 molar solution and ammonium acetate. The diastereomer is the first of both diastereomers to leave the column.

¹ H-NMR spectrum (500 MHz, D ₂ 0, 2 rotamers in approximately 5: 1 ratio): δ 0.74 (m, minor rotamer), 1.01 (m,

2H), 1.10 to 1.33 (m, 3H), 1.48 to 1.88 (m, 15H, thereof 1.51 (d, 3H)), 1.92 to 2.12 (m, 3H), 1.96 (s, acetate), 2.30 (m,

1H), 3.20 (m, 3H), 3.38 (m, 1H), 3.47 (q, minor rotamer), 3.53 to 3.68 (m, 2H), 3.73 (m, 1H), 4.20 (d, minor rotamer), 4.33 (m, IH), 4.38 (m, IH), 4.51 (d, minor rotamer) ppm.

1 H-NMR (125 MHz, D ₂ O): guanidine: δ 157.38, carbonyl carbons, δ 174.11, 173.45 and 168.64 ppm.

Example 9

Production method of HOOC- (R or S) CH (Me) - (R) Cha-Pro-Agm / b x HOAc

The diastereomer that emerges from the column after the first diastereomers in the separation of Example 8 is the title compound named above.

1 H-NMR spectrum (500 MHz, D ₂ O, 2 rotamers in approximately 9: 1 ratio): δ 0.88 (m, minor rotamer), 1.05 (m,

2H), 1.12 to 1.33 (m, 3H), 1.42 (broad s, 1H), 1.50 to 1.88 (m, 15H, thereof 1.55 (d, 3H)), 1.93-2.13 (m, 3H), 1.95 (s, acetate), 2.30 (m, 1H), 2.40 (m, minor rotamer), 3.22 (t, 2H), 3.28 (t, 2H), 3.64 (m, 1H), 3.70 (q, 1H), 3.98 (t, minor rotamer), 4.35 (t, 1H), 4.41 (t, 1H); dd, 1H) ppm.

Example 10

HOOC- (R) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm / x HOAc

Alkylation as in Example 4, using H- (R) Cha-Pro-Agm (Z) (see Example 3) and Br-CH ( ⁿ Pr) COOEt and method (e) deprotection followed by method ( b) to give the compound

HOQC- (R, S) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm. The title compound is obtained by separating the diastereomers by reverse phase high performance liquid chromatography, eluting with acetonitrile and 0.1 molar ammonium acetate solution (1: 4). and lyophilizing the water after evaporating the solvent. This diastereomer is the first of both columns.

1 H-NMR spectrum (300 MHz, MeOD): δ 0.8-1.1 (m, 5H, of which 0.92 (t, 3H)), 1.1-1.1 (m, 22H), 1 95 (s, acetate), 2.2 (m, 1H), 3.1-3.35 (m, 5H), 3.48 (m, 1H), 3.88 (m,

1H), 4.0 (m, 1H), 4.4 (dd, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.50, carbonyl carbons: δ 168.55 and 174.16 ppm.

Example 11

HOOC- (R) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm / b x HOAc

Another diastereomer from separation in Example 10, which starts from the column after the first diastereomer, is the title compound named above.

¹ H-NMR spectrum (500MHz, MeOD) : δ 0.85 to 1, , 05 (m, 5H of which 0.95 (t, 3H); 2.08 (m, 22H), 1.9 (with, acetate), 2.14 (m, 1H), 3.1 to 3, 4 (m, 5H) , 3.45 (m, IH), 3.62 (m, 1H); 3.80 (m, 1H), 4.34 (dd, IH) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.53, carbonyl carbons: δ 169.01 and 174.27 ppm.

Example 12

Production method of HOOC- (R or S) CH (Ph) - (R) Cha-Pro-Agm / bx HOAc (i) Production method ^t BuOOC- (R or S) CH (Ph) - (R) Cha-Pro- Agm (Z)

A mixture of 0.55 mmol of H- (R) Cha-Pro-Agm (Z) (see Example 3), 0.66 mmol of tert-butyl (R, S) phenyl (bromoacetate) and 1.4 mmol of potassium carbonate in 10 ml of acetonitrile was stirred at room temperature for 28 hours and then for a further 5 hours at 60 ° C. The diastereomeric mixture (3: 1 by NMR spectrum) was filtered and evaporated. The remaining oil was twice subjected to flash chromatography, eluting with dichloromethane: methanol 92: 8. The result is a complete separation of the two diastereomers [(R _f = 0.36 (minor isomer) and 0.27 (major isomer)].

1 H-NMR spectrum of major isomer (500.13 MHz, CDCl ₃ ): δ 0.79 (q, 1H), 0.90 (q, 1H), 1.06 to 1.70 (m, H), 1 37 (s, 9H), 1.85 to 2.03 (m, 3H), 2.20 (m, 1H), 3.10 to 3.24 (m, 3H), 3.25 to 3.38 (m, 2H), 3.42 (m, 1H), 3.53 (m, 1H), 4.30 (s, 1H), 4.49 (dd, 1H), 5.08 (s, 2H) 7.19-7.40 (m, 10H); broad NH signals are observed in the 6.7-8.6 ppm range.

(ii) Production method of HOOC- (R or S) CH (Ph) - (R) Cha-Pro-Agm / bx HOAc mmol of major isomer and 0.5 mmol of thioanisole are dissolved in trifluoroacetic acid and kept at room temperature for 8 hours. hours. After evaporation at 13 Pa over 5 hours, the residual oil is purified by reverse-phase high performance liquid chromatography, eluting with a 2: 2 mixture of acetonitrile and 0 M ammonium acetate:

3. The title compound is obtained after evaporation of the solvent and lyophilization.

¹ H-NMR spectrum (500.13 MHz, MeOD): δ 0,85 to 1.01 (m, 2H), 1 13 to 1 38 (m, 4H), 1.53 to 2.05 (m, 14H), 1.92 (s, acetate) , 2.18 (m, 1H) 3.08 to 3.26 (m, 3H) , 3.32 to 3.45 (m,

2H), 3.64 (m, 1H), 3.93 (t, 1H), 4.37 (dd, 1H), 4.43 (s, 1H), 7.28-7.50 (m, 5H) ) ppm.

^P3 C-NMR (125.6 MHz, MeOD): guanidine: [delta] 158.7, carbonyl carbons: [delta] 173.8, 174.7, 177.0 ppm.

Example 13

Production Method of HOOC- (R, S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm x HOAc

Alkylated as in Example 4, using H- (R) Cha-Pro-Agm (Z) (see Example 3) and

Br-CH (CH ₂ -CH ₂ -Ph) -COOEt and method (a) for deprotection followed by process (e) to give HOOC- (R, S) CH (CH ₂ -CH ₂ - Ph) - (R) Cha-Pro-Agm.

Example 14

Production method of HOOC- (R or S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm / ax 2 TFA

The title compound is obtained by separating the diastereomers obtained in Example 13 by reverse phase high performance liquid chromatography, eluting with a 2: 3 mixture of acetonitrile and 0.1 molar ammonium acetate, and lyophilizing water and trifluoroacetic acid, after evaporation of the solvent. This diastereomer, which is the first of the two diastereomers of the column, is the title compound.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.93-1.11 (m, 2H), 1.24 (m, 1H), 1.29-1.40 (m, 2H), 52 to 1.85 (m,

11H), 1.89-2.11 (m, 4H), 2.14-2.32 (m, 3H), 2.83 (t, 2H), 3.14 (t, 2H), 3.24 (t, 2H), 3.50 (q, 1H), 3.70 (m, 1H), 4.00 (t, 1H), 4.36-4.42 (m, 2H), 7.17-7 7.31 (m, 5H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.66, carbonyl carbons: δ 168.08, 171.53, 174.16 ppm.

Example 15

Method for producing HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Agm (H) (i) Method for producing BnOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Agm (Z)

1.1 equivalents of benzyl acrylate and 1 equivalent of H- (R) Cha-Pro-Agm (Z) (see Example 3) were dissolved in ethanol (20 mL / mmol) and stirred at room temperature for 20 hours. The solvent was evaporated and the crude material was purified by flash chromatography, eluting with a 95: 5 mixture of dichloromethane and methanol saturated with ammonia. Finally, the solvent was evaporated and the material was dried under reduced pressure.

¹ H-NMR spectrum (500 MHz, CDCl3) ): δ 0.7 to 0.95 (m, 2H), 1, 0 to 1 1.5 (m, 10H), 1.5-1.75 (m, 5H), 1.75 to 1.92 (m, 2H) , 2.0 | m, 1H), 2.17 (broad s, 1H) 2.45 (m, 2H) 2.63 (m (1H), 2.79 (m, 1H), 2.97-3, 25 (m, 4H) 3.33 (m, 2H), 3, 52 (broad t, 1H); 4.45 (broad d, 1H), 4.95-7 5.12

(m, 4H), 7.13-7.4 (m, 10H) ppm.

(ii) Production method of HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Agm x HOAc

The compound is produced by applying process (a) of deprotection to the product (i) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 0.88 (m, 2H), 1.00 to 1.23 (m, 3H), 1.33 (broad s, 1H), 1.42 to 1.72 (m, 11H), 1.78 to 2.00 (m, 3H), 1.94 (s, acetate), 2.18 (m, 1H), 2.52 (m, 2H) 3.03-3.20 (m, 6H), 3.50 (m, 1H), 3.72 (m, 1H), 4.23 (m, 1H), 4.30 (m, 1H) ppm .

13,

C-NMR (125 MHz, D ₂ O): guanidine: carbonyl carbons: δ 178.07, 173.96, 168.24 ppm.

δ 157,25,

Example 16

Process for the production of EtOOC-CO- (R) for Cha-Pro-Agm (H) (i) Process for the production of EtOOC-CO- (R) for Cha-Pro-Agm (Z)

To a cold solution of 0.46 g (0.89 mmol)

H- (R) Cha-Pro-Agm (Z) (see Example 3) and 199 mg (1.97 mmol) of N-methylmorpholine in 10 mL of -10 ° C tetrahydrofuran were added 134 mg (0.98 mmol). Cl-COCOOEt dissolved in 3 mL of tetrahydrofuran. The mixture was maintained at -10 ° C for 1 hour and then stirred at room temperature for an additional hour. The solvent was evaporated and the residue was dissolved in ethyl acetate. The organic phase was washed twice with water and dried over sodium sulfate. Evaporation of the solvent and crystallization from ethyl acetate gave 0.275 g of the title compound as white crystals. Yield: 50%.

(ii) A method for producing EtOOC-CO- (R) Cha-Pro-Agm x HOAc

The compound is produced by applying process (b) of deprotection to the product (i) above.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.9 to 2.25 (m,

¢.,.

24H, thereof 1.17 (t, 3H)), 1.90 (s, acetate), 3.1 to 3.25 (m, 4H), 3.5 to 3.65 (m, 3H, thereof 3.59 (q, 2H)), 3.88 (m, 1H), 4.35 (m, 1H), 4.69 (dd, 1H) ppm.

¹³ C-NMR (75.5 MHz, MeOD): guanidine: δ 157.56, carbonyl carbons: δ 159.21, 160.74, 172.81, 174.56 ppm.

Example 17

Production method of (R, S) Bia- (R) Cha-Pro-Agm x 2 TFA

It is alkylated as in Example 4, using H- (R) Cha-Pro-Agm (Z) (see Example 3) and α-bromobutyrolactone followed by deprotection according to method (a). The title compound is obtained as a mixture of 2 diastereomers. 1 H-NMR spectrum (500 MHz, D ₂ O, mixture of diastereomers)

in a ratio of about 1: 1): δ 0,93 to 1,06 (m, 2H), 1.09 to 1.30 (m 3H), 1,37 to 1.49 (m, IH), 1.50 to 1.87 (m, 11H), 1.8 to 9 2.10 (m, 3H) , 2.24 to 2.36 (m, 1H), 2.44 to 2.56 (m, IH); 2.72 to 2.85 (rn, 1H), 3.10 up to 3.30 (m, 4H), 3.56 to 3.65 (m, 1H), 3,75 to 3.84 (m, IH); 4.2 to 5.0 (m, 5H) , partial

covered by H-O-D signal) ppm.

¹³ C-NMR (125.76 MHz, D ₂ 0): guanidine: δ 157.34 (peaks overlapping); carbonyl carbons: δ 174.34, 173.90, 173.62, 167.88, 167, 58 (2 peaks overlap) ppm.

Example 18

Production method of HOOC- (R or S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm / bx 2 TFA

The title compound is obtained by treating the diastereomer of Example 13 in the same manner as described in Example 14. This diastereomer exits from the first diastereomer.

1 H-NMR spectrum (500 MHz, MeOD): δ 0.95-1.06 (m, 2H), 1.14-1.40 (m, 4H), 1.48-1.84 (m, 11H), 1.87-2.30 (m, 6H), 2.72-2.90 (m, 2H), 3.12-3.32 (m, 4H),

3.52 (m, 1 H), 3.72 (m, 1 H), 4.04 (dd, 1 H), 4.27 (t, 1 H),

4.37 (dd, 1H), 7.17-7.32 (m, 5H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.68, carbonyl carbons: δ 168.14, 171.46, 174.03 ppm.

Example 19

H- (R) Cha-Pro-Nag x 2 HOAc (i) Z- (R) Cha-Pro-NH- _{(CH2) 3} -NH (Boc)

To a solution of 1 mmol of Z- (R) Cha-PrO-OSu in 1 mL of dimethylformamide at 0 ° C is added H ₂ N- (CH ₂ ) ₃ -NH (Boc) (see production of starting materials) which is dissolved in 1 ml of dimethylformamide and the pH was adjusted to about 9 by addition of N-methylmorpholine. The reaction mixture was stirred at room temperature for about 3 days and then poured onto water. The aqueous phase is extracted four times with ethyl acetate. The combined organic phases were washed twice with 0.3 molar potassium hydrogen sulphate solution, 0.2 molar sodium hydroxide solution, brine and dried. Evaporation and flash chromatography, eluting with 4: 1 ethyl acetate: petroleum ether gave the title compound. Yield: 59%.

(ii) A method for producing Z- (R) Cha-Pro-NH- (CH ₂ ) ₃ -NH ₂

0.6 mmol of Z- (R) Cha-Pro-NH- _{(CH2) 3} -NH (Boc) was dissolved in 8 ml of dichloromethane. To the solution was added 2 mL of trifluoroacetic acid, and the reaction mixture was stirred for 1 hour. The solvent was evaporated and the residue was dissolved in dichloromethane, washed twice with 0.2 M sodium hydroxide solution and dried over sodium sulfate. Evaporation of the solvent gave the amine. Yield: 93%.

¹ H-NMR spectrum (500 MHz, CDC1 _3): δ 0.79 to 1.03 (m, 2H), 1.05 to 10.6 1.75 (m, 15H); 1.84 to 2.08 (m, 4H), 2.36 (m, 1H), 2.66 (m, 2H), 3.25 (m, 2H), 3.43 (q, 1 H), 3.85 (m, 1H), 4.45 (m, IH), 4.56 (d, 1 H), 5.09 (m, 2H); 5.35 (d, 1H); 7.30

up to 7.45 (m, 5H) ppm.

(iii) A method for producing Z- (R) Cha-Pro-Nag x HOAc

0.55 mmol (1 equivalent) of Z- (R) Cha-Pro-NH- (CH ₂ ) ₃ -NH ₂ was dissolved in 2 mL of dimethylformamide and adjusted to pH 8-9 with triethylamine. equivalent) of 3,5-dimethyl-1-pyrazolylformamidinium nitrate dissolved in 1 ml of dimethylformamide was added to the solution and the reaction mixture was stirred at room temperature for 3 days. The solvent was evaporated, the crude material was lyophilized to remove water, and purified by reverse-phase high performance liquid chromatography, eluting with a 4: 6 mixture of acetonitrile and 0.1 molar ammonium acetate to give the title compound after evaporation of the solvent. lyophilization to remove water. Yield: 93%. 1 (iv) Production method of H- (R) Cha-Pro-Nag x 2 HOAc

The compound is produced by applying deprotection according to method (a) on. the product (iii) above.

i '-'!

^{± 1} H-NMR spectrum (500 MHz, D ₂ 0): δ 0.82 to 1.03 (m,

2H), 1.03 to 1.28 (m, 3H), 1.35 (m, 1H), 1.53-1 , 82 (m, 9H), 1,82 to 2.05 (m, 3H), 1.89 (s, acetate), 2.24 (m, ÍH), 3.15 (t, 2H) , 3.23 (q, 2H) 3.55 (m, 1H), 3.72 (m, 1H) , 4.27 to 4 , 34 m 2H) ppm.

¹³ C-NMR (75.5 MHz, D ₂ O): guanidine: δ 157.37, carbonyl carbons: δ 169.81, 174.52 ppm.

Example 20

Production method ⁿ Bu- (R) Cha-Pro-Nag x 2 HOAc (i) Production method H- (R) Cha-Pro-Nag (Z)

The compound was prepared from Boc- (R) Cha-Pro-OSu and Boc-Nag (Z) using the same method as described for H- (R) Cha-Pro-Agm (Z) in Example 3.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.8-1.03 (m,

2H), 1.10 to 1.50 (m, 6H) 1.60 to 1.83 (m, 8H), 1.87 to 2.20 (m, 3H), 3.15 (m, 1 H), 3.25 (m, [0100] 2H ), 3.42 (m , 2H) , 3.63 (dd, 1H), 3.70 (m, 1H), 4.36 (broad s, 1H, 5.07 (WITH, 2H), 7.22-7 7.43 (m, 5H) ppm. (ϋ) Method v production ⁿ Bu- (R) Cha-Pro- Nag (Z)

0.5 g (1 mmol) of H- (R) Cha-Pro-Nag (Z) is dissolved in 10 ml of methanol. To the solution was added 0.1 g (1 mmol) of triethylammonium hydrochloride, 44 mg (0.7 mmol) of sodium cyanoborohydride, followed by 76 mg (1.05 mmol) of butyraldehyde, and the reaction mixture was stirred at room temperature for 20 hours. The solvent was evaporated and the crude material was dissolved in ethyl acetate, washed twice with water, once with brine and dried over sodium sulfate. The solvent was evaporated and the crude material was purified by flash chromatography, eluting with ethyl acetate / ethanol / triethylamine 88: 10: 2. Finally, the solvent was evaporated and the material was dried under reduced pressure. Thus, 0.22 g ^{of n} Bu- (R) Cha-Pro-Nag (Z). Yield: 40%.

1 H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.82 to 1.0 (m, 5H, of which 0.88 (t, 3H)), 1.08 to 1.49 (m, 10H), 1.58 to 1.8 (m, 7H), 1.88 to 2.22 (m, 3H), 2.4 (m, 1H), 2.5 (m, 1H),

3.05 (broad s, 1H), 3.3 (m, 1H), 3.4-3.53 (m, 3H), 3.73 (m, 1H), 4.42 (broad s, 1H) 5.1 (s, 2H), 7.25-7.43 (m, 5H) ppm.

(iii) Production method ⁿ Bu- (R) Cha-Pro-Nag x 2 HOAc

The compound is produced by deprotection when applying process (a) to product (ii) above.

1 1 H-NMR spectrum (300MHz, d ₂ 0) : δ to 1 , 31 (m, 3H) 1.38 (m, 3H) 1 , 55 and ž 1, 2.15 (m, 3H), 1.95 (s, acetate), 2.34 (m, 3.08 (m, 1H), 3.24 (t, 2H), 3.30 (m, 2H), (m, 1H), 4.32 (t, 1 H), 4.41 (dd, 1H) ppm.

0.94 (t, 2H); (m, 11H); 1H); 2.95 (m,

3.66 (m, IH)

1.10 1.88 to

1H), 3.82 ¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.40, carbonyl carbons: δ 180.39, 174.28, 168.55 ppm.

Example 21

Production method of HO- (CH ₂ ) ₃ - (R) Cha-Pro-Nag x 2 TFA (i) Production method of BnO- (CH ₂ ) ₃ - (R) Cha-Pro-Nag (Z)

0.5 g (1 mmol) of 1-benzyloxy- (3-trifluoromethanesulfonylpropane) (see preparation of starting materials) and H- (R) Cha-Pro-Nag (Z) (see Example 20) are dissolved in 10 ml of tetrahydrofuran.

To the reaction mixture was added 0.28 g (2 mmol) of potassium carbonate and stirred at room temperature for 2 hours. The solvent was evaporated and the crude material was extracted with a mixture of ethyl acetate and water. The organic phase was washed once with aqueous sodium bicarbonate solution, once with water and once with brine. After drying over sodium sulfate, the solvent is evaporated and the crude material is subjected to flash chromatography, eluting with a 95: 5 mixture of dichloromethane and methanol saturated with ammonia. Finally, the solvent is evaporated and the substance is dried under reduced pressure. 0.29 g of the title compound is obtained. Yield: 45%.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.77-1.03 (m,

2H), 1.03-2.18 (m, 19H), 2.52 (m, 1H), 2.64 (m, 1H), 3.03 (broad s, 1H), 3.1 to 3, Δ (m, 7H), 3.66 (m, 1H), 4.41 (broad s, 1H), 4.46 (s, 2H), 5.08 (s, 2H), 7.2-7, 4 (m,

5H), 7.55 (m, 1H) ppm.

(ii) A method for producing HO- (CH ₂ ) ₃ - (R) Cha-Pro-Nag x 2 TFA

The compound is produced by applying deprotection method (a) to the product (i) above.

1 H-NMR spectrum (500 MHz, D ₂ O): δ 1.00 (broad s,

2H), 1.10 to 1.32 (m, 3H), 1.40 (broad s, 1H), 1.55 to 2.15 (m, 14H), 2.30 (m, 1H), 3, 05-3.35 (m, 6H), 3.57-3.75 (m, 3H), 3.81 (broad s, 1H), 4.35 (broad s, 1H), 4.42 (broad s (1H) ppm.

Example 22

Production method of HOOC-CH ₂ '(R) Cha-Pro-Nag x HOAc (i) Production method of H- (R.) Cha-Pro-NH- (CH ₂ ) ₃ - N ₃

The compound was prepared in the same manner as H- (R) Cha-Pro-Agm (Z) (see Example 3), starting from Boc- (R) Cha-Pro-OSu and Boc-NH- (CH ₂ ) ₃ -. N ₃ , which replaces Boc-Agm (Z).

(ii) Production method of EtOOC-CH ₂ - (R) Cha-Pro-NH- (CH ₂ ) ₃ - NH ₂ x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-NH- (CH ₂ ) ₃ -N ₃ and EtOOC-CH ₂ -Br followed by deprotection by method (a) to reduce the azide yields the title compound. compound.

(iii) Production method of EtOOC-CH ₂ - (R) Cha-Pro-Nag x HOAc

The same method as described in Example 19 (iii) for Z- (R) Cha-Pro-Nag was used to achieve the amine guanidation of part (ii) above. The title compound was removed in pure form after reverse phase high performance liquid chromatography, eluting with a mixture of acetonitrile and 0.1 molar acetate. Ammonium in ratio 3:

7, evaporating the solvent and removed water by lyophilization.

(iv) Production method of HOOC-CH ₂ - (R) Cha-Pro-Nag x HOAc

The compound is produced by applying method (e) of deprotection to the product (iii) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 0.99 (m, 2H), 1.09

to 1.30 (m, 3H), 1.44 (m, 1H) , 1.59 to 2 09 (m, 12H), 1.92 (s, acetate), 2.29 (m, 1H) 20 (t, 2H), 3.28 (m, 2H), 3.52 to 3.63 (m, 3H), 3.76 (m, 1H) , 4.38 (dd, 1H), 4.42 (t, IH) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.43, carbonyl carbons: δ 168.72, 171.36, 174.35 ppm.

Example 23

EtOOC-CH ₂ - (R) Cha-Pro-Nag x HOAc

The compound was prepared according to Example 22 (iii).

1 H-NMR spectrum (300 MHz, D ₂ O): δ 1.07 (m, 2H) 1.17 to 1 59 (m, 7H, thereof) 1, 38 (t, 3H) 2.24 (m, 12H) 2.04 (s, acetate), 2.39 (m, 1H), 3.31 (t, 2H), 3.39 (t, 2H), 3.63 up to 3.90 (m, 4H), 4.12 (t, 1H), 4.36 (q, 2H), 4.46 (dd,

1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.37, carbonyl carbons: δ 173.73, 175.09, 175.70 ppm.

Example 24

Production Method ¹ PrOOC-CH ₂ - (R) Cha-Pro-Nag x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and Br-CH ₂ COcApr followed by deprotection by method (b) affords the title compound.

1 H-NMR spectrum (500 2H), 1.1-2.15 (m, 22H, z)

3H), 1.92 (s, acetate), 2.2

3.4 (m, 1H); 3.55 (m, 1H);

1H), 5.03 (m, 1H) ppm.

MHz, MeOD) : δ 0.85 up to 1.05 (m, of which 1.23 (d, 3H), 1.25 (d, (m, 1 H), 3 , 10 to 3 35 (m, 5 H), , 65-3.8 (m, . 2H) , 4.28 (dd,

¹³ C-NMR (125 MHz, D ₂ O) carbonyl carbons: δ 170.40, 172.00 and guanidine: δ 174.50 ppm.

157,39,

Example 25

A process for producing ^t _BuOOC-CH2 - (R) Cha-Pro-Nag x 2 TFA

Alkylation as in Example 4 using H- (R) Cha-ProNag (Z) (see Example 20) and Br-CH ₂ CO 3 ^t Bu followed by deprotection by method (b) affords the title compound.

1 H-NMR spectrum (300 MHz, MeOD): δ 0.9 to 1.15 (m, 2H), 1.15 to 2.15 (m, 25H, thereof 1.55 (br s, 9H)) 2.3 (m, 1H), 3.15-3.45 (m, 4H), 3.55 (m, 1H), 3.7-3.95 (m, 3H), 4.3-4 4 (m, 2H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.55, carbonyl carbons: δ 166.55, 168.13 and 174.33 ppm.

Example 26

Production method H00C-CH ₂ -00C-CH ₂ - (R) Cha-Pro-Nag x HOAc (i) Production method BnOOC-CH ₂ -OOC-CH ₂ - (R) Cha-Pro-Nag (Z)

0.20 g (0.40 mmol) of H- (R) Cha-Pro-Nag (Z) (see Example 20) is mixed with 0.115 g (0.40 mmol) of benzyloxycarbonylmethyl bromoacetate, 55 mg (0.40 mmol) potassium carbonate and 5 ml of acetonitrile. The mixture was stirred at room temperature for 6 hours. The solvent was evaporated and the crude material was chromatographed, eluting with a 9: 1 mixture of dichloromethane and methanol. 0.20 g of the desired compound is obtained after evaporation of the solvent. Yield: 71%.

(ii) Production method of HOOC-CH ₂ -OOC-CH ₂ - (R) Cha-Pro-Nag x HOAc

The compound is produced by applying process (a) of deprotection to the product (i) above.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.85-1.1 (m, 2H), 1.1-1.6 (m, 8H), 1.6-1.15 (m, 10H) 1.99 (s, acetate), 2.23 (m, 1H), 3.1 to 3.4 (m, 4H), 3.45 to 3.65 (m, 4H), 3.7 to 3.4 3.9 (m, 3H), 4.34 (m, 1H), 4.48 (dd, 2H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.8, carbonyl carbons: δ 176.1, 175.2, 174.9, 173.1 ppm.

Example 27

Production Method of H ₂ N-CO-CH ₂ - (R) Cha-Pro-Nag x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and C1-CH ₂ CONH ₂ in the presence of a catalytic amount (10 mol%) of potassium iodide in the reaction, followed by deprotection group (a) provides the title compound

-NMR spectrum (500 MHz, D ₂ O): δ 1.2 (m, 2H), 1 , 12 a 1.34 (m, 3H), 1.46 (m, IH), 1.61 up to 2.13 (m, 9H), 1.99 (s, acetate), 2.34 (m, 1H) 3, 25 (t, 2H), 3.33 (t, 2H) , 3.60 and ž 3.82 (m, 4H), 4.22 (t, IH), 4, 41 (dd, 1H) ppm. 13 C-NMR spectrum (75 MHz, D ₂ O) guanidine: δ 157, 5,

carbonyl carbons: δ 168.94, 169.40, 174.43 ppm.

Example 28

Production method of HOOC-CH ₂ -NH-CO-CH ₂ - (R) Cha-Pro-Nag x 2 TFA

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (See Example 20) and Br-CH ₂ CONHCH ₂ -COOBn (See Preparation of starting materials) followed by deprotection procedure (a) gave the title compound.

1 H-NMR spectrum up to 1.38 (m, 1H), 3.83 (m, 4.45 (m, (m, 3H), 1.47 3.17 to 3.26 IH), 3.88 ( s, 2H) ppm.

(500 MHz, MeOD): δ 1.01 (m, 2H), 1.15 (m, 1H), 1.64 to 2.13 (m, 12Η), 2.27 (m, 3Η), 3, 37 (m, 1H); 3.51 (m, 1H);

2H), 3.93 to 4.06 (m, 2H), 4.35 to ¹³ C-NMR (75 MHz, MeOD): guanidine: δ 158.71 and carbonyl carbons: δ 166.94, 168.35 , 172.44, 174.17 ppm.

Example 29

A method for producing (HOOC-CH _{2) 2} ^_ (R) Cha-Pro-Nag x HOAc (i) Preparation of _{(EtOOC-CH2) 2-} (R) Cha-Pro-NH- (CH _{2) 3} -NH ₂ x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-NH- (CH ₂ ) ₃ -N ₃ (see Example 22) and Br-CH ₂ COOEt (10 equivalents used for alkylation) followed by deprotection by method (a) affords the title compound.

(ii) Production method of (EtOOC-CH ₂ ) ₂ - (R) Cha-Pro-Nag x HOAc

The same method as described in Example 19 (iii) for Z- (R) Cha-Pro-Nag was used to perform the guanidation of the amine obtained above. Purification was accomplished by reverse phase high performance liquid chromatography using acetonitrile and a 0.1 molar ammonium acetate solution of 4: 6 to elute.

(iii) Production method of (HOOC-CH ₂ ) ₂ - (R) Cha-Pro-Nag x HOAc

Hydrolysis of the ester group is carried out by the deprotection method described in (e) using twice the amount of sodium hydroxide. The final compound was purified by reverse phase high performance liquid chromatography using a 2: 8 mixture of acetonitrile and 0.1 molar ammonium acetate to elute, evaporate the solvent and remove the water by lyophilization.

1 H-NMR spectrum (300 MHz, D ₂ O): δ 0.92-1.49 (m,

6H), 1.60-2.54 (m, 10H), 2.05 (s, acetate), 3.25-3.50 (m, 4H), 3.65-4.03 (m, 6H, of which 3.95 (s, 4H)), 4.49 (m,

1 H), 4.71 (m, 1 H, partially overlapped by H-O-D peak) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.64, carbonyl carbons: δ 168.62, 171.39, 174.30 ppm.

Example 30

Production method of HOOC-CH ₂ - (Me) - (R) Cha-Pro-Nag x 2 TFA (i) Production method of Me- (R) Cha-Pro-Nag (Z)

The compound was prepared from Boc- (Me) - (R) Cha-Pro-OSu and Boc-Nag (Z) in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3.

(ii) Method for producing HOOC-CH ₂ - (Me) - (R) Cha-Pro-Nag x 2 TFA

Alkylation as in Example 4 using Me- (R) Cha-Pro-Nag (Z) and Br-CH ₂ COOBn followed by deprotection by method (b) affords the title compound.

1 H-NMR spectrum (500 MHz, D ₂ O): δ 0.8-1.06 (m,

2H), 1.08-1.27 (m, 4H), 1.55-2.10 (m, 12H), 2.30 (m,

1H), 3.04 (s, 3H), 3.14-3.33 (m, 4H), 3.63 (m, 1H), 3.81

(m, 1H), 4.13 (apparent broad s, 2H), 4.38 (broad dd, 1H),

4.56 (broad t, 1H) ppm.

¹³ C-NMR (125.76 MHz, D ₂ O): guanidine: δ 157.40, carbonyl carbons: δ 174.05, 168.83, 167.44 ppm.

Example 31

Production method of HOOC-CH ₂ - ( ⁿ Bu) - (R) Cha-Pro-Nag X 2 TFA

Alkylation as in Example 4 using ⁿ Bu- (R) Cha-Pro-Nag (Z) (see Example 20) and Br-CH ₂ COOBn followed by deprotection by method (a) affords the title compound.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 0.78-0.88 (m,

3H), 0.88 to 1,02 (m, 2H), 1.02 to 1.23 (m, 4H). 1.23 to 1.38 (m, 2H), 1.45 to 1.84 (m, 11H) , 1.84-2.10 (m, 3H) 2.24 (m, IH), 3.05 to 3.18 (m, 3H) 3.18 to 3.38 (m, 3H) 3.57 (m, IH), 3.77 (m,  IH), , 4.05 to 4.25 (m, 2H). 4.32 (m, IH), 4.50 (m, IH) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 159.17 carbonyl carbons: δ 175.66, 171.13, 169.31 ppm.

Example 32

Production method of HOOC- (R, S) CH (Me) - (R) Cha-Pro-Nag x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and Br-CH (Me) COOBn followed by deprotection by method (a) affords the title compound as a mixture of two diastereomers.

Example 33

72 Production Method of HOOC- (R or S) CH (Me) - (R) Cha-Pro-Nag / a x HOAc

The compound is obtained by resolution of the diastereomers formed in Example 32 using reverse phase high performance liquid chromatography, eluting with acetonitrile and 0.1 molar ammonium acetate 1: 4, followed by evaporation of the solvent. The diastereomer exits the column first as the two diastereomers.

¹ H-NMR spectrum (300 MHz, D ₂ O, 2 rotamers in ratio)

approximately 9: 1): δ 0.78 (m, minority rotamer), 1.07 (m, 2H), 1.17 to 1.42 (m, 3H), 1.48-1.64 (m, 4H, z of it 1.56 (d, 3H)) 1,64 to 1.95 (m, 9H) 2.20 (m, 3H), 2.00 (s, acetate), 2.37 (m, 1H); 3.28 (t, 2H) 3.38 (t, 2H), 3.53

(m, minor rotamer), 3.63 (m, 2H), 3.77 (m, 1H), 4.24 (d, minor rotamer), 4.35-4.50 (m, 2H), 4, 60 (d, minor rotamer) ppm.

Example 34

Production method of HOOC- (R or S) CH (Me) - (R) Cha-Pro-Nag / b x HOAc

The title compound is obtained by applying the same method as described in Example 33 to the compound formed in Example 32. This diastereomer is eluted from the column after the first diastereomer.

1 H-NMR spectrum (300 MHz, D ₂ O, 2 rotamers in ratio)

approximately 9: 1) δ 0, 95 ( m, minority rotamer), 1.12 (m, 2H), 1.22 to 1 , 40 (m, 3H), 1.40 to 1.67 (m, 4H, thereof 1.60 (d, 3H)) 1.67 to 2.00 (m, 9H), 2.00 to 2.00 2.25 (m, 3H). 2.03 (s, acetate ), 2 , 40 (m, 1H), 3.25 to 3.48 (m, 4H) 3.66 and ž

3.84 (m, 2H), 3.93 (m, 1H), 4.38 (m, 1H), 4.50 (m, 1H), 4.93 (m, minor rotamer) ppm.

¹³ C-NMR (75.5 MHz, D ₂ O): guanidine: δ 157.42, carbonyl carbons: δ 168.05, 171.99, 174.04 ppm.

Example 35

Method for producing EtOOC- (R, S) CH (Me) - (R) Cha-Pro-Nag x 2 TFA

The compound was prepared in the same manner as described in Example 22 using EtOOC-CH (Me) -Br instead of Br-CH ₂ COOEt in alkylation.

¹ H-NMR spectrum (500 MHz, MeOD, 2 diastereomers in a ratio of approximately 2.5: 1 as well as 4 rotamers): δ 0.88 to 2.43 (m, 25H), 3.1 to 4.55 (m (11H) ppm.

1 H-NMR (75 MHz, MeOD): guanidine: δ 158.65, carbonyl carbons: δ 174.33, 170.66, 168.20 ppm.

Example 36

Process for the manufacture of HOOC- (R or S) CH ( ⁿ Pr) - (R) Cha-Pro-Nag / ax HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and Br-CH ( ⁿ Pr) COOEt and deprotection by method (e) and then method (e) affords HOOC - (R, S) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm. The title compound is obtained by separating the diastereomers (this diastereomer elutes as the first of two diastereomers from the column) using reverse phase high performance liquid chromatography, eluting with a 1: 4 mixture of acetonitrile and 0.1 molar ammonium acetate, evaporation of the solvent and removal of water by lyophilization.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.85-1.05 (m,

5H, of which 0.95 (t, 3H)), 1.1 to 2.05 (m, 20H), 1.95 (s, acetate), 2.18 (m, 1H), 3.15 to 3 3 (m, 4H); 3.35 (m, 1H);

3.46 (m, 1H), 3.85 (m, 1H), 4.04 (m, 1H), 4.38 (dd, 1H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.73, carbonyl carbons: δ 171.63, 174.43 and 176.78 ppm.

Example 37

Production method of HOOC- (R) CH (CH ₂ -OH) - (R) Cha-Pro-Nag x 2 TFA

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) a

Br- (S) CH (CH ₂ -Obn) -COOBn followed by deprotection by method (a) to give the title compound.

¹ H-NMR spectrum (300 MHz, D ₂ O): δ 0.75 to 1.56 (m, 7H), 1.56 to 2.30 (m, 11H), 2.40 (m, 1H), 3.15 to 3.55 (m, 4H), 3.55 to 4.60 (m, 7H) ppm.

Example 38

Production method of HOOC- (R, S) CH (Ph) - (R) Cha-Pro-Nag x 2 TFA

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and Br-CH (Ph) COO-Bu and deprotecting according to method (a) and then method (f) to give the title compound as a mixture of 2 diastereomers.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.8-1.1 (m,

2H), 1.1-2.18 (m, 16H), 2.26 (m, 1H), 3.04-3.35 (m, 5H), 3.45 (m, 1H), 3.7 (m, 1H), 4.35 (m, 1H), 4.85 (s, 1H, one isomer), 5.05 (s, 1H, second isomer), 7.4-7.6 (m, 5H) 7.75 (broad t, 1H) ppm.

¹³ C-NMR (75 MHz, MeOD): guanidine: δ 158.68, carbonyl carbons: δ 174.39, 174.15 and 170.5, 170.06 and 168.32, 167.78 ppm.

Example 39

Production method HOOC- (S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Nag x HOAc

Alkylation as in Example 21 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and TfO- (R) CH (CH ₂ CH ₂ Ph) -COOEt and deprotecting according to method (e) and then by method (a) affords the title compound.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.77-1.05 (m, 2H), 1.05-1.35 (m, 5H), 1.35-2.16 (m, 14H) 1.87 (s, acetate), 2.71 (t, 2H), 3.07-3.53 (m, 7H), 3.73 (m, 1H),

4.32 (m, 1H), 7.03-7.25 (m, 5H) ppm.

¹³ C-NMR (75 MHz, MeOD): guanidine: δ 158.71, carbonyl carbons: δ 174.15, 177.31, 182.61 ppm.

Example 40

Production method HOOC- (R) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Nag x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and Br-CH (CH ₂ CH ₂ Ph) COOEt followed by deprotection by method (a) and then method (e) provides the compound

HOOC- (R, S) CH (CH ₂ -CH ₂ -Ph) - (R) Cha-Pro-Nag. The title compound is obtained by separating the diastereomers by reverse phase high performance liquid chromatography, using acetonitrile and a 0.1 molar ammonium acetate solution (2: 3) to elute and lyophilize the water after evaporation of the solvent.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.97 (m, 2H), 1.10

to 1, 41 m 3H), 1 , 43 to 2, , 30 (m, 16H), 1.96 (s, acetate), 2.70 (m, 2H) , 3.06 to 3.26 (m, (3H), 3.28-3.66 (m, 3H) 3.84 (m, lH) , 4.14 (wide t, IH), 4.39 (dd, IH) , 7.11 to 7.28 (m, 5H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 158.66 ppm.

Example 41

Method for the production of HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Nag x HOAc (i) Method for the production of EtOOC-CH ₂ -CH ₂ - (R) Cha-Pro-NH- (CH ₂ ) ₃ - NH ₂

Alkylation as described in Example 15, using H- (R) Cha-Pro-NH- (CH ₂ ) 3 -N ₃ instead of H- (R) Cha-Pro-Agm (Z) followed by deprotection by ( a), provides the title compound.

(ii) _{EtOOC-CH2 -CH2} - (R) Cha-Pro-Nag x HOAc

Guanidation of the amine listed above in the same manner as described in Example 19 for Z- (R) Cha-Pro-Nag affords the title compound (ii).

(iii) Production method of HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Nag x HOAc

The compound is produced using method (e)

to remove the protecting group on the product (ii) mentioned above. ¹ H-NMR spectrum (500 MHz, D ₂ 0): δ 1.12 (m, 2H) , 1.22 to 1.48 (m, 3H), 1.54 (broad s, 1H), 1.70 to 2.37 (m, 12H), 2.14 (s , acetate), 2.53 (m, 1H), 2.70 (broad s, 2H ), 3 , 15 (t IH), 3 25 to 3.55 (m, 5H), 3.75 (m, 1H), 3.93 (m, IH), 4.43

(t, 1H), 4.52 (m, 1H) ppm.

Example 42

EtOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Nag x HOAc

The compound was prepared according to Example 41 (ii).

1 H-NMR spectrum (500 MHz, D ₂ O): δ 0.97 (m, 2H), 1.11

to 1.39 (m, 7H) , of which 1.30 (t,  3H)) , i, 50 (t , 2H) , 1.61 to 1.76 (m, 5H), 1.76 to 2.14 (m, 5H), 1.93 (with, acetates t), 2.29 (m, 1H), 2.62 (t, 2H) 2.77 to 2.94 (m, 2H), 3, 23 (t, 2H) 3.32 (t, 2H), 3.60 to 3.87 (m, 3H), 4.20 (q, 2H), 4.36 (dd, 1H) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.39, carbonyl carbons: δ 182.05, 175.13, 175.02 ppm.

Example 43

Method for producing HOOC- (CH ₂ ) ₃ - (R) Cha-Pro-Nag x HOAc (i) Method for producing EtOOC-CH = CH-CH ₂ - (R) Cha-Pro-Nag (Z) equivalent to H- (R) Cha-Pro-Nag (Z) (see Example 20) and 1.1 equivalents of ethyl 3-bromocrotonate are dissolved in 15 mL / mmol of acetonitrile. Potassium carbonate was added to the reaction mixture and stirred at room temperature for 2 hours. After filtration and evaporation of the solvent, the crude material was purified by flash chromatography, eluting with a mixture of dichloromethane and methanol. Finally, the solvent was evaporated and the compound was dried under reduced pressure.

1 H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.73-1.0 (m,

2H), 1.0 to 1.4 (m, 8H, of which 1.33 (t, 3H)), 1.43 to 2.15 (m, 12H), 2.96 (broad s, 1H), 3.12 (dd, 1H), 3.16-3.48 (m, 6H), 3.56 (m, 1H), 4.15 (q, 2H), 4.35 (broad s, 1H),

5.03 (s, 1H), 6.0 (d, 1H), 6.85 (dt, 1H), 7.05 (broad s,

1H), 7.17-7.37 (m, 5H), 7.5 (broad s, 1H) ppm.

(ii) Method for producing EtOOC- (CH ₂ ) 3 - (R) Cha-Pro-Nag x 2 TFA

The compound is produced by applying process (a) of deprotection to the product (i) above.

(iii) Production method of HOOC- (CH ₂ ) 3 - (R) Cha-Pro-Nag x 2 HOAc

The compound is produced by applying method (e) of deprotection to the product (ii) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 1.02 (broad s,

2H), 1.08-1.32 (m, 3H), 1.42 (broad s, 1H), 1.55-2.15 (m, 14H), 1.92 (s, acetate), 2, 33 (br s, 3H), 3.00 (br s, 1H), 3.07 (br s, 1H), 3.18-3.40 (m, 4H), 3.62 (br s, 1H) 3.82 (broad s, 1H), 4.33 (broad s, 1H),

4.40 (broad s, 1H) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.42, carbonyl carbons: δ 181.87, 174.34, 168.64 ppm.

Example 44

Production method of EtOOC- (CH ₂ ) 3 - (R) Cha-Pro-Nag x 2 TFA

The compound was prepared according to Example 43 (ii).

¹ H-NMR spectrum (300 MHz, MeOD / D ₂ O): δ 0.63-1.30 (m, 9H, thereof 1.02 (t, 3H)), 1.30-1.97 (m 14H), 2.06 (broad s, 1H), 2.28 (m, 2H), 2.72-3.20 (m, 6H), 3.36 (m, 1H), 3.60 (m 1 H, 3.94 (m, 2 H), 4.06 (m, 1 H), 4.17 (m, 1 H)

- 79 ppm.

¹³ C-NMR (75 MHz, MeOD / D ₂ O): guanidine: δ 158.10, carbonyl carbons: δ 175.40, 174.23, 168.54 ppm.

Example 45

Production method of HOOC-CO- (R) Cha-Pro-Nag x HOAc (i) Production method of EtOOC-CO- (R) Cha-Pro-Nag (Z)

0.50 g (0.97 mmol) of H- (R) Cha-Pro-Nag (Z) was dissolved in 0.54 ml of triethylamine and 8 ml of dichloromethane. To the reaction mixture was added 0.146 g (1.07 mmol) of ethyloxalyl chloride dissolved in 2 mL of dichloromethane, increasing the temperature from 22 to 28 ° C. The reaction mixture was then stirred at room temperature for 2 hours. The organic phase is washed twice with water, dried over sodium sulphate and subjected to flash chromatography, eluting with a mixture of ethyl acetate and 99% ethanol (9: 1).

1. 92 mg of the title compound are obtained. Yield: 15%.

(ii) A method for producing HOOC-CO- (R) Cha-Pro-Nag x HOAc

Application of the deprotection method (b) to the product and then method (e) affords the title compound.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.88-1.14 (m, 2H), 1.15-1.5 (m, 4H), 1.5-2.3 (m, 13H) 1.9 (s, acetate), 3.1 to 3.43 (m, 4H), 3.6 (m, 1H), 4.05 (m, 1H),

4.43 (dd, 1H), 4.5 (m, 1H) ppm.

¹² C-NMR (75 MHz, D ₂ O): guanidine: δ 157.57, carbonyl carbons: δ 165.94, 173.95, 174.85 and 181.22 ppm.

Example 46

Production method of MeOOC-CO- (R) Cha-Pro-Nag x HOAc (i) Production method of MeOOC-CO- (R) Cha-Pro-Nag (Z)

The methyl ester is obtained by transesterification of EtOOC-CO- (R) Cha-Pro-Nag (Z) (see Example 45) in a flash chromatography column using a 9: 1 mixture of ethyl acetate and methanol as the eluent. .

(ii) A method for producing MeOOC-CO- (R) Cha-Pro-Nag x HOAc

The compound is produced by applying method (b) of deprotection to the product (i) above.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.9-1.1 (m, 2H), 1.1-2.3 (m, 17H), 1.9 (s, acetate), 3, 12 to 3.4 (m,

4H), 3.52-3.67 (m, 2H), 3.9 (s, 3H), 4.35 (m, 1H), 4.65 (m, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.52, carbonyl carbons: δ 159.11, 161.20, 173.17 and 174.90 ppm.

Example 47

Production method of (R, S) Bia- (R) Cha-Pro-Nag x 2 TFA

It is alkylated as in Example 4 using H- (R) Cha-Pro-Nag (Z) (see Example 20) and α-bromobutyrolactone followed by deprotection by method (a). The title compound is obtained as a mixture of 2 diastereomers.

- '• H-NMR (300 MHz, D ₂ 0, mixture of diastereomers):

δ 1.0 to 1.43 (m, 5H), 1.45 to 1.60 (broad s, 1H), 1.64 to 1.60,

2.28 (m, 12H), 2.31-2.50 (m, 1H), 2.80-2.98 (m, 1H),

3.23 to 3.46 (m, 4H), 3.66 to 3.79 (m, 1H), 3.82 to 3.96 (m,

1H), 4.33-5.08 (m, 5H, partially obscured by H-O-D signal) ppm.

Example 48

Production method H00C- (R, S) CH (CH ₂ COOH) - (R) Cha-Pro-Nag x HOAc (i) Production method BnOOC- (R, S) CH (CH ₂ COOBn) - (R) Cha- Pro-Nag

0.21 g (0.42 mmol) of H- (R) Cha-Pro-Nag (Z) (see Example

20) and 0.12 g (0.42 mmol) of dibenzyl maleate are dissolved in 10 ml of acetonitrile. The mixture is refluxed overnight and flash chromatographed, eluting with dichloromethane / methanol (94: 6). Evaporation of the solvent affords the title compound. Yield: 22%.

(ii) Production method of HOOC- (R, S) CH (CH ₂ COOH) - (R) Cha-Pro-Nag x HOAc

The compound is produced by applying method (a) of deprotection to the product (i) above.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.9 to 2.4 (m, 19H), 2.00 (s, acetate), 2.7 to 3.0 (m, 2H), 3, 1 to 3.6 (m,

5H), 3.75-3.9 (m, 2H), 4.2-4.5 (m, 2H) ppm.

Example 49

Production method of MeOOC- (R, S) CH (CH ₂ COOMe) - (R) Cha-Pro-Nag x HOAc (i) Production method of MeOOC- (R, S) CH (CH ₂ COOMe) - (R) Cha- Pro-Nag

0.21 g (0.42 mmol) of H- (R) Cha-Pro-Nag (Z) (see Example

20) and 0.24 g (1.7 mmol) of dimethyl maleate are dissolved in 15 ml of methanol. The mixture was refluxed overnight, evaporated and subjected to flash chromatography, eluting with a 9: 1 mixture of dichloromethane and methanol. Evaporation of the solvent gave the title compound. Yield: 45%.

(ii) Production method of MeOOC- (R, S) CH (CH ₂ COOMe) - (R) Cha-Pro-Nag x HOAc

The compound is produced by applying method (c) of deprotection to the product (i) above.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.85-1.1 (m,

2H), 1.15-2.3 (m, 17H), 1.91 (s, acetate), 2.6-2.8 (m, 2H), 3.1-3.5 (m, 5H) 3.5 to 3.8 (m, 10H, of which 4 singlets 3.66, 3.68, 3.71, 3.73), 4.29 (m, 1H) ppm.

Example 50

Production method of HOOC-Ph-4-CH ₂ - (R) Cha-Pro-Nag x 2 TFA (i) Production method ^υΒ-ΡΗ-4-ΟΗ ₂ - (R) Cha-Pro-NH- (CH ₂ ) ₃ ~ N ₃

0.39 g (1.1 mmol) H- (R) Cha-Pro-NH- _{(CH2) 3-N3} (See Example 22) and 0.33 g (1.2 mmol) of tert-butyl p-bromomethylbenzoate was dissolved in 10 ml of acetonitrile and 0.19 g (2.4 mmol) of potassium carbonate was added. The mixture was refluxed overnight and evaporated. The crude material was subjected to flash chromatography, eluting with a 92: 8 mixture of dichloromethane and methanol to give 0.50 g of the title compound. Yield 84%.

(ii) Production method ^t BuOOC-Ph-4-CH ₂ - (R) Cha-Pro-NH- (CH ₂ ) ₃ -NH ₂

To a solution of 0.60 g (1.8 mmol) of bis-phenylthiostannane,

0.20 g (1.8 mmol) of thiophenol and 0.18 g (1.8 mmol) triethylamine in 50 ml dichloromethane at 0 ° C was added 0.50 g (0.92 mmol) ^t BuOOC-Ph-4 -CH ₂ - (R) Cha-Pro-NH- (CH ₂ ) ₃ -N ₃ . The mixture was stirred at 0 ° C for 30 minutes and at room temperature for 4 hours. The reaction mixture was then diluted with dichloromethane and washed with aqueous sodium bicarbonate solution, and washed successively three times with 2% hydrogen peroxide. The organic layer was extracted with dilute hydrochloric acid. The combined acidic aqueous phases were washed with ethyl acetate and then basified with aqueous sodium hydroxide solution. The aqueous layer was extracted twice with ethyl acetate. The combined organic layer was dried over sodium sulfate and evaporated. Flash chromatography, eluting with an 8: 2 mixture of dichloromethane and methanol saturated with ammonia gave 0.12 g of the title compound. Yield: 26%.

(iii) Production method of HOOC-Ph-4-CH ₂ - (R) Cha-Pro-Nag x 2 TFA

Guanidation of the amine above in the same manner as described in Example 19 for Z- (R) Cha-Pro-Nag, followed by deprotection by method (f), affords the title compound.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0,9 to 1,5 (m, 7H), 1,4 to 1, 9 (m, 9H) 2.1 (m (2H), 2.16 (m , IH), 2.32 (m, 1H) 3.2 to 3.3 (m, 3H) , 3.41 (pentet, IH) 3.53 (m IH), 3.77 (m, 1H), 4.2 to 4.3 (m , 3H), 4.42 (dd, 1H), 7.15 (d, 2H), 8.10 (d, 2H) ppm. ¹³ c-nmr : spectrum (125 MHz , MeOD) : guanidine: δ 160,8,

carbonyl carbons: δ 174.3, 168.9 and.168.2 ppm.

Example 51

(HO) ₂ P (O) -CH ₂ - (R) Cha-Pro-Nag x HOAc mg (92 mmol) (EtO) ₂ PO-CH ₂ - (R) Cha-Pro-Nag (Z) ( see Example 53) is dissolved in 3 ml of acetonitrile. 0.15 ml of trimethylsilyl bromide was added to the reaction mixture and left at room temperature for 21 hours. Evaporation and NMR spectral analysis revealed some ester remaining. The crude material was redissolved in 3 mL of acetonitrile and 0.15 mL of trimethylsilyl bromide was added. After 5 hours, evaporate and purify by reverse phase high performance liquid chromatography, eluting with 30:70 acetonitrile: 0.1 molar ammonium acetate. After filtration, evaporation and lyophilization, the title compound is obtained. Yield: 8%.

@ ³ H-NMR spectrum (500 MHz, MeOD): δ 0.8 up to 1.1 (m, 2H), 1.15 to 1.15 1.4 (m, 4 H), 1, , 5 to 1.9 (m, 10H) 1.9 to 2.1 (m, 4H), 1.96 (s, acetate), 2.20 (m, 1H), 2.95 (m, 1H), 3.0 to 3.0 3.2 (m, 3 H), 3.4 to 3.5 (m, 2H), 4.09 (m, IH) , 4.39 (broad

d, 1H), 4.59 (m, 1H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.6, carbonyl carbons: δ..174, 2 and 170.6 ppm.

Example 52

Production method Et0 (H0) P (O) -CH ₂ - (R) Cha-Pro-Nag (Z) x 2 HOAc (i) Production method (EtO) (HO) P0-CH ₂ - (R) Cha-Pro -Nag (Z) mg (77 mmol) of (EtO) ₂ PO-CH ₂ - (R) Cha-Pro-Nag (Z) (see Example 53) is dissolved in 2 mL. ethanol and 2 ml of 2M sodium hydroxide solution. The mixture was stirred overnight and evaporated.

The crude material was purified by reverse phase high performance liquid chromatography, eluting with a 30:70 mixture of acetonitrile and 0.1 molar ammonium acetate. After filtration and evaporation of the solvent, the title compound was obtained.

(ii) Production method (EtO) (HO) PO-CH ₂ - (R) Cha-Pro-Nag x 2 HOAc

The compound is produced by applying method (c) of deprotection to the product (i) above.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.9-1.1 (m,

2H), 1.15-1.35 (m, 6H, of which 1.28 (t, 3H)), 1.35-1.5 (m, 2H), 1.5-1.6 (m, 1H), 1.65-1.8 (m, 6H), 1.9-2.1 (m, 3H), 1.95 (s, acetate), 2.19 (m, 1H), 2.8 to 3.0 (m, 2H), 3.1 to 3.25 (m, 2H), 3.27 (m, 1H), 3.36 (m, 1H), 3.48 (m, 2H)

1H), 3.9-4.05 (m, 4H), 4.36 (broad d, 1H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.6, carbonyl carbons: δ 175.0 and 174.7 ppm.

Example 53

Production method (EtO) ₂ P (O) -CH ₂ - (R) Cha-Pro-Nag x HOAc (i) Production method (EtO) ₂ PO-CH ₂ - (R) Cha-Pro-Nag (Z)

0.2 g (0.40 mmol) of H- (R) Cha-Pro-Nag (Z) (see Example 20) was dissolved in 5 mL of tetrahydrofuran and 0.11 g (0.80 mmol) of potassium carbonate was added to the solution. and 0.12 g (0.40 mmol) of diethyl triflylmethylphosphonate (triflate refers to the trifluoromethanesulfonic acid salt). The mixture was stirred at room temperature for 2 hours. The reaction mixture was treated with water and the aqueous layer was extracted three times with ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated. It will

I with 0.14 g of the title compound. Yield: 53%.

(ii) Production method of (EtO) ₂ PO-CH ₂ - (R) Cha-Pro-Nag x HOAc

The compound is produced by applying process (c) of deprotection to the product (i) above.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.85-1.05 (m, 2H), 1.15-1.3 (m, 5H), 1.34 (t, 6H), 5 to 1.85 (m, 8H), 1.9 to 2.05 (m, 3H), 1.91 (s, acetate), 2.10 (m, 1H), 2.22 (m, 1H) 2.90 (dd, 1H), 3.05 (dd, IH), 3.1-3.3 (m, 3H), 3.42 (m, IH), 3.53 (m, IH), 3.71 (dd, 1H), 3.82 (m, 1H), 4.1 to 4.2 (m, 4H), 4.28 (dd, 1H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.7, carbonyl carbons: δ 176.0 and 175.1 ppm.

Example 54

Production method of HOOC-CH ₂ - (R) Cha-Pro-Mag x HOAc (i) Production method of H- (R) Cha-Pro-NH- (CH ₂ ) ₂ -NH (Z)

The compound is prepared from Boc- (R) Cha-Pro-OSu and NH ₂ - (CH ₂ ) ₂ - NH (Z) in the same manner as described for H- (R) Cha-Pro-Agm (Z) in the example 3.

(ii) Production method of EtOOC-CH ₂ - (R) Cha-Pro-NH- (CH ₂ ) ₂ -NH ₂ x HOAc

Carry out the alkylation as in Example 4 followed by deprotection according to method (a) to give the title compound.

(iii) Production method of HOOC-CH.?-( R)Cha-Pro-Mag x HOAc

Guanidation of the amine above in the same manner as described in Example 19 for Z- (R) Cha-Pro-Nag, followed by deprotection by method (e), affords the title compound after purification by reverse-phase high performance liquid chromatography. elution with a mixture of acetonitrile and a 0.1 molar ammonium acetate solution and removal of water by lyophilization.

2H), 10H) 3.50 4.54 1 H-NMR spectrum (300 MHz, D ₂ O): δ 1.19 to 1.43 (m, 3H), 1.52 (m, 1H), 2 0.06 (s, acetate), 2.31 to 2.47 (m, (m, 2H), 3.60 to 3.75 (m, 3H), 3.85 (m, 2H) ppm.

0.90 to 1 1.63 to 2 IH), 3.44 (m, IH),

(m, (m, (m, 2H), from 46 to ¹³ C-NMR (75 MHz, D ₂ 0): guanidine: δ 157.82 and carbonyl carbons: δ 168.80, 171.41, 174, 81 ppm.

Example 55

Production method of H- (R, S) Pro (3-Ph) -Pro-Agm x 2 TFA

The compound was prepared from Boc- (R, S) Pro (3-Ph) -Pro-OSu (see Production of starting materials) in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3 , followed by deprotection by method (b).

¹ H-NMR spectrum (300 MHz, D ₂ O, mixture of 2 diastereomers with unknown relative stereochemistry): δ 1.0 to 1.8 (m, 7H),

2.0 to 2.5 (m, 3H), 2.8 to 4.3 (m, 10H), 4.56 (d, 1H, major rotamer), 4.90 (d, 1H, major rotamer), 7.2 to 7.5 (m, 5Η) ppm.

¹³ C-NMR (125.76 Hz ^, D ₂ 0): guanidine: δ 157.36 (minor and major), carbonyl carbons:

δ 174.1 (major rotamer), 174.0 (minor rotamer),

167.8 (major rotamer), 167.0 (minor rotamer) ppm.

Example 56

Production method of H- (R, S) Pro (3- (trans) Ch) -Pro-Agm x 2 TFA

This compound was prepared from Boc- (R, S) Pro (3- (trans) Ch) -Pro-OSu (see production of starting compounds) according to the method described for H- (R) Cha-Pro-Agm (Z) in the example 3, followed by deprotection by method (b).

1 H-NMR spectrum (500 MHz, D ₂ O, 1.8: 1 mixture of 2 diastereomers): δ 0.95 to 1.32 (m, 5H), 1.35 to 1.46 (m (1H), 1.50-1.92 (m, 10H), 1.93-2.15 (m, 4H), 2.23-2.43 (m, 2H), 3.15-3.30 (m, 4H), 3.35-3.50 (m, 2H), 3.57-3.68 (m, 1H), 3.74-3.82 (ra, 1H), 4.34-4 41 (m, 1H), 4.51 (d, 1H, minor rotamer), 4.48 (d, 1H, major rotamer) ppm.

¹³ C-NMR (125.76 MHz, D ₂ 0): guanidine: δ 157.36 (minor and major), carbonyl carbons: δ 174.34 (major), 174.07 (minor), 168 , 94 (minor and major rotamer) ppm.

Example 57

HOOC-CH ₂ - (R, S) Pro (3- (trans) Ph) -Pro-Agm x 2 TFA (i) H- (R, S) Pro (3- (trans) Ph) - Pro-Agm

The compound is prepared from Boc- (R, S) Pro (3- (trans) Ph) -Pro-QSu (see production of starting compounds) in the same manner as described for the compound

H- (R) Cha-Pro-Agm (Z) in Example 3.

(ii) Production method of HOOC-CH ₂ - (R, S) Pro (3- (trans) Ph) -Pro-Agm x TFA

It is alkylated as in Example 4 using Br-CH ₂ COOBn followed by deprotection by method (b). The title compound is obtained as a mixture of 2 diastereomers.

¹ H-NMR spectrum (500 MHz, MeOD, mixture of 2 diastereomers 1.1: 1): δ 1.40 to 1.80 (m, 6H), 1.85 to 2.05 (m, 1H), 2.10 to 2.30 (m, 1H), 2.50 to 2.65 (m, 2H), 3.10 to 3.40 (m, 6H), 3.50 to 3.70 (m, 2H) 3.9 to 4.40 (m, 4H),

4.63 (d, 1H, major rotamer), 4.67 (d, 1H, minor rotamer), 7.30-7.60 (m, 5H) ppm.

¹³ C-NMR (125.76 MHz, D ₂ 0): guanidine: δ 157.52 (both isomers), carbonyl carbons: δ 173.87, 173.73, 169.12, 168.94, 167.21 and 167.00 ppm.

Example 58

HOOC-CH ₂ - (R, S) Pro (3- (trans) Ph) -Pro-Nag x 2 TFA (i) Production Method of H- (R, S) Pro (3- (trans) Ph) - Pro-Nag

Compound is prepared from Boc- (R, S) Pro (3- (trans) Ph) -Pro-OSu (see production of starting compounds) and Boc-Nag (Z) according to the same procedure as described for H- (R) Cha-Pro-Agm (Z) in Example 3.

(ii) Production method of HOOC-CH ₂ - (R, S) Pro (3- (trans) Ph) -Pro-Nag x TFA

It is alkylated as in Example 4 using Br-CH ₂ COOBn followed by deprotection according to method (b). The title compound is obtained as a mixture of 2 diastereomers.

1 H-NMR spectrum (500 MHz, MeOD, mixture of 2 diastereomers in a ratio of approximately 1.5: 1): δ 1.40 to 1.85 (m, 4H), 1.90 to 2.00 (m, 1H ), 2.10-2.30 (m, 1H), 2.45-2.70 (m, 2H), 3.08-3.46 (m, 6H), 3.57-3.70 (m 2H, 3.90-4.0 (m, 1H), 4.32-4.40 (m, 1H), 4.04 and 4.29 (AB-quartet, 2H, major rotamer), 4, 16 and 4.37 (AB-quartet, 2H, minor rotamer), 4.60 (d, 1H, major rotamer), 4.64 (d, 1H, minor rotamer), 7.3-7.6 (m, 5H) ppm.

¹³ C-NMR (125.76 MHz, D ₂ 0): guanidine: δ 157.48 (both isomers), carbonyl carbons: δ 173.90, 173.71, 169.01, 168.94 and 167.07 (both isomers) ppm.

Example 59

Production method of HOOC-CH ₂ - (R) Cha-Pic-Agm x 2 TFA (i) Production method of H- (R) Cha-Pic-Agm (Z)

The compound was prepared from Boc- (R) Cha-Pic-OSu (see production of starting compounds) according to the same method as described for H- (R) Cha-Pro-Agm (Z) in Example 3.

(ii) A method for producing HOOC-CH ₂ - (R) Cha-Pic-Agm x 2 TFA

It is alkylated as in Example 4 using Br-CH ₂ COOBn followed by deprotection by method (a). This gives the title compound.

@ ¹ H-NMR (75 MHz, MeOD): .delta. 1.2 (m, 2H), 1.13-1

2.00 (m, 20H), 2.24 (broad d, 1H), 3.12-3.45 (m, 5H),

3.71 (broad d, 1H), 3.87 (s, 2H), 4.65 (broad t, 1H), 5.06 (m, 1H) ppm.

1 H-NMR (75 MHz, D ₂ O): guanidine: δ 157.47, carbonyl carbons: δ 169.42, 170.03 and 172.71 ppm.

Example 60

Production method of HOOC-CH ₂ - (Me) - (R) Cha- (R, S) Pic-Agm x HOAc (i) Production method of Me- (R) Cha- (R, S) Pic-Agm (Z)

The compound was prepared from Boc- (Me) - (R) Cha-Pic-OSu in the same manner as described for H- (R) Cha-Pro-Agm (Z) in the example.

(ii) _HOOC-CH2 - (Me) - (R) Cha- (R, S) Pic-Agm x HOAc

It is alkylated as in Example 4 using Br-CH ₂ COOBn followed by deprotection by method (b). The title compound is obtained.

Note: To some extent during synthesis, epimerization of S-pipecolinic acid occurs.

^{The 1} H-NMR spectrum consists of a complex consisting of 2 diastereomers in a ratio of approximately 1: 1 and their rotamers.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.75 to 2.15 (several m, 20H), 1.95 (broad s, acetate), 2.2 to 2.7 (6H, observed 2 distant signal sets at a ratio of about 1: 1, of which 2.35 and 2.55 (s, 3H)), 3.0 to 3.5 (m, 6H), 3.9 to 4.17 (m, 2H) of which 4.14 (dd)), 4.4 to 4.5 (m, 1H), 4.97 to 5.15 (two broad dd, 1H) ppm.

C-NMR (75 MHz, D 2 O): guanidine: δ 157.50, carbonyl carbons: δ 169.65, 170.01, 170.54, 172.67 and 172.89 ppm.

Example 61

Production method of HOOC- (R, S) CH (Me) - (R) Cha-Pic-Agm x TFA

Alkylation as in Example 4 using H- (R) Cha-Pro-Agm (Z) (see Example 59) and Br-CH (Me) COOBn followed by deprotection by method (a). The title compound is obtained as a mixture of two diastereomers.

Example 62

Production method of HOOC- (R or S) CH (Me) - (R) Cha-Pic-Ag / a x 2 TFA

The compound is obtained by separating the diastereomers of Example 61 using reverse phase high performance liquid chromatography, eluting with a 1: 3 mixture of acetonitrile and 0.1 molar ammonium acetate, followed by evaporation of the solvent and removal of water and trifluoroacetic acid by lyophilization. . This diastereomer is eluted first in both diastereomers from the column.

³ H-NMR spectrum (300 MHz, D ₂ 0, ² rotamers in approximately 5: 1 ratio): δ 0.70 (m, minor rotamer), 0.75 to 1.0 (m, 2H), 1.0 to 1.28 (m, 3H), 1.28 to 1.83 (m, 20H) thereof

1.57 (d, 3H)), 2.14 (broad d, 1H), 2.92 (t, minor rotamer), 3.03-3.32 (m, 5H), 3.59 (broad d, 1H), 3.85 (q, minor rotamer), 3.98 (q, IH), 4.30-4.50 (m, minor rotamer), 4.54 (m, IH), 4.95 (s) (1H) ppm.

³³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.39, carbonyl carbons: δ 172.26 (2 carbons) and 169.92 ppm.

Example 63

Production method of HOOC- (R or S) CH (Me) - (R) Cha-Pic-Agm / b x TFA

The title compound was obtained using the same procedure as described in Example 62 for the compound formed in Example 61. This diastereomer was eluted first in the column.

¹ H-NMR spectrum (500 MHz, D ₂ O, 2 rotamers in approximately 5: 1 ratio): δ 0.72 (m, minor rotamer), 0.82 (m, minor rotamer), 0.97 (m, 2H), 1.0 to 1.23 (m, 3H), 1.23 to 1.40 (m, 2H), 1.40 to 1.83 (m, 18H, thereof 1.63 (d, 3H) 1.11, 2.11 (d, 1H), 2.17 (d, minor rotamer), 2.92 (t, minor rotamer), 3.05-3.25 (m, 4H), 3.29 (t (IH), 3.74 (d, IH), 4.02 (q, IH), 4.34 (d, minor rotamer), 4.41 (dd, minor rotamer), 4.52 (t, IH) , 4.95 (s, 1H) ppm.

13,

C-NMR (125 MHz, D ₂ 0): guanidine: δ 154.68 and carbonyl carbons: δ 169.81, 169.60, 167.36.

Example 64

Production method of HOOC-CH ₂ -CH ₂ - (R) Cha-Pic-Agm x 2 TFA

The compound was prepared from H- (R) Cha-Pro-Agm (Z) (see Example 59) in the same manner as described for HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Agm in Example 15 with except that 1.2 equivalents of benzyl acrylate is used instead of 1.1 equivalents.

^b H-NMR spectrum (500 MHz, D ₂ O, 2 rotamers in approximately 4: 1 ratio): δ 0.70 to 0.90 (m, minor rotamer), 0.90 to 1.0 (m, 2H) 1.05-1.25 (m, 3H), 1.39-1.45 (m, 2H), 1.45-1.85 (m, 15H), 2.1 (broad d, 1H), 2.2 (wide

d, minor rotamer), 2.75 (t, 2H), 2.95 (t, minor rotamer), 3.1 to 3.4 (m, 7H), 3.75 (broad d, 1H), 4, 55 (t, 1 H), 4.95 (m, 1 H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.48, carbonyl carbons: δ 170.10, 172.58 and 174.75 ppm.

Example 65

Production method of H- (R) Cha-Pic-Nag x 2 TFA (i) Production method of Boc- (R) Cha-Pic-Nag (Z) (i) The compound is prepared from Boc- (R) Cha-Pic-OSu. in the same manner as described for Boc- (R) Cha-Pro-Agm (Z) in the example.

(ib) A production method starting from Boc- (R) Cha-Pic-OH

0.432 mL (2 mmol) of diphenylphosphoryl azide was added to a stirred solution of 765 mg (2 mmol) of Boc- (R) Cha-Pic-OH in 5 mL of dimethylformamide at -10 ° C. After 10 minutes, 600 mg (2.1 mmol) of H-Nag (Z) x 2 HCl (see preparation of starting compounds) in 5 ml of tetrahydrofuran and 615 mg (4.4 mmol) of triethylamine are added. The reaction mixture is kept on an ice bath for 3 hours and then at room temperature. at room temperature for 12 hours and then poured into water. Extraction of the aqueous phase with ethyl acetate followed by drying of the organic phase with magnesium sulphate and evaporation of the solvent under reduced pressure gave 1.18 g of a substance consisting of a mixture of diastereomers (acid epimers)

S-pipecoline) in a ratio of 97: 3 (RS / YY). Yield 96%.

(ic) A production method starting from, Boc- (R) Cha-Pic-OH

EDC hydrochloride (4.2 g, 21.9 mmol) was added to a stirred solution of EDC hydrochloride (8 g, 20.9 mmol) at -15 ° C.

Boc- (R) Cha-Pic-OH, 10.6 g (88 mmol) of N, N-dimethylaminopyridine and 6.3 g (19.5 mmol) of H-Nag (Z) x 2 HCl (see production of starting compounds) in acetonitrile. The reaction mixture was allowed to warm to +15 ° C over 16 hours. The solvent was evaporated under reduced pressure and the residue was dissolved in ethyl acetate.

Washing with water, 0.3 molar potassium bisulfite solution, 0.3 molar sodium bicarbonate, water and brine, followed by drying over sodium sulfate and evaporation of the solvent gave 11.9 g of a mixture of diastereomers (acid epimers)

S-pipecoline) in a ratio of 98: 2 (RS / RR). Yield: 92.5%.

¹ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.85 to 2.0 (m,

29H, of which 1.40 (broad s, 9H)), 2.46 (broad d, 1H), 3.1 to 11.5

3.4 (m, 5H), 3.92 (broad d, 1H), 4.53 (broad q, 1H), 5.10 (s, 2H), 5.22 (broad s, 1H), 5, 29 (broad d, 1H), 6.7 to 11.2

7.2 (broad, 3H), 7.25 to 7.45 (m, 5H) ppm.

¹³ C-NMR (125 MHz, CDCl ₃ ): guanidine: δ 156.9, carbonyl carbons: δ 173.6, 170.3, 163.7 and 161.7 ppm.

(ii) Production method of H- (R) Cha-Pic-Nag (Z)

The compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3, starting from Boc- (R) Cha-Pic-Nag (Z).

1 H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.8-2.0 (m, 22H), 2.24 (broad d, 1H), 3.1-3.4 (m, 5H), 3.72 (broad d, 1H), 3.84 (broad q, 1H), 5.05 (broad d, 1H), 5.08 (s,

2H), 7.3-7.5 (m, 5H) ppm.

(iii) A method for producing H- (R) Cha-Pic-Nag x 2 TFA

The compound is produced by applying process (a) deprotection to the product (ii) produced above.

1 H-NMR spectrum (500 MHz, MeOD): δ 0.9-1.1 (m,

2H), 1.2-2.0 (m, 18H), 2.32 (broad d, 1H), 3.20 (t, 2H), 3.30 (t, 2H), 3.36 (m, 1H), 3.69 (broad d, 1H), 4.49 (dd,

1H), 5.05 (broad d, 1H) ppm.

¹³ C-NMR (125 MHz, MeOD): guanidine: δ 158.7, carbonyl carbons: δ 172.7 and 171.4 ppm.

Example 66

Method for the production of Me- (R) Cha- (R, S) Pic-Nag x 2 TFA (i) Method for the production of Me- (R) Cha- (R, S) Pic-Nag (Z)

The compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3, starting from Boc- (Me) - (R) Cha-Pic-OSu and Boc-Nag (Z). ). Acid epimerization

The S-pipecoline takes place during the synthesis and the substance is obtained as a mixture of two diastereomers.

(ii) A method for producing Me- (R) Cha- (R, S) Pic-Nag x 2 TFA

The compound is produced by applying method (b) of deprotection.

^{The 1} H-NMR spectrum is a complex consisting of 2 diastereomers with a ratio of about 4: 1 and their rotamers.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.8-1.08 (m,

2H), 1.15 to 2.4 (several m, 19H), 2.6 to 2.75 and 2.9 to 2.95 (...

- 97 (several s, 3H), 3.1 to 3.6 (several m, 5H), 3.75 to 4.1 (several m, IH), 4.4 to 4.7 (several m, IH) , 5.05 to 5.15 (two dd, 1H) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 154.84, carbonyl carbons: δ 167.60 and 169.99 ppm.

Example 67

Process for producing HOOC-CH ₂ - (R) Cha-Pic-Nag (i) Process for producing BnOOC-CH ₂ - (R) Cha-Pic-Nag (Z)

Alkylation as in Example 4 using H- (R) Cha-Pic-Nag (Z) (see Example 65) and Br-CH ₂ COOBn. The title compound is obtained.

⁴ H-NMR spectrum (500 MHz, CDCl ₃ ): δ 0.8-1.0 (m, 2H), 1.1-1.7 (m, 19H), 1.79 (broad d, 1H), 2.3 to 2.5 (m 2H, of which 2.38 (broad d, 1H)), 3.00 (broad t, 1H), 3.1 to 2.5

3.4 (m, 5H, of which 3.38 (d, 1H)), 3.58 (d, 1H), 3.6-3.7 (m, 2H), 5.06 (dd, 2H), 5 , 07 (s, 2H), 5.16 (broad s, 1H),

6.7 to 7.1 (broad signal, 1H), 7.15 (broad s, 1H), 7.2 to 7.1

7.4 (m, 10H) ppm.

¹³ C-NMR (125 MHz, _CDCl3): guanidine and carbonyl carbons: δ 176.0, 173.6, 170.8, 163.8 and 161.7.

(iia) Process for producing HOOC-CH ₂ - (R) Cha-Pic-Nag x 2 HCl

Removal of the protecting group by method (a) followed by purification by reverse phase high performance liquid chromatography, eluting with a 1: 3 mixture of acetonitrile and 0.1 molar ammonium acetate, by evaporation at 40 to 50 ° C. ° C and lyophilization affords the title compound as acetate. Treatment with a 20-fold excess of hydrochloric acid, evaporation and re-lyophilization affords the bis-hydrochloride of the title compound.

1 H-NMR spectrum (500 MHz, D ₂ O, mixture of 2 rotamers): δ 0.7 to 2.0 (m, 20H), 2.17 (broad d, 1H), 2.95 (t, minor rotamer) 3.17 (t, 2H), 3.25-3.35 (m, 3H), 3.72 (broad d,

1H), 3.86 (dd, minor rotamer), 3.90 (s, 2H), 4.72 (t,

1H, 4.99 (broad s, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine δ 157.4, carbonyl carbons: δ 169.9, 170.2 and 173.0 ppm.

(iib) Production method of HOOC-CH ₂ - (R) Cha-Pic-Nag x 2 HBr

BnOOC-CH ₂ - (R) Cha-Pic-Nag (Z) is dissolved in a 95: 5 mixture of isopropyl alcohol and water and hydrogenated to 5% palladium on carbon at atmospheric pressure in the presence of 2.2 equivalents of hydrobromic acid. The catalyst was filtered off and the solvent was evaporated. A yellow oil is obtained. (Alternatively, the acid may be added after hydrogenation and filtration.) Crystallization from isopropanol (or ethanol) and ethyl acetate 1: 1 gives the title compound as a white crystalline powder.

¹ H-NMR spectrum (500 MHz, D ₂ O, mixture of 2 rotamers): δ 1.15 to 2.0 (m, 20H), 2.30 (broad d, 1H), 3.30 (m, 2H) 3.40 to 3.50 (m, 3H), 3.85 to 3.90 (m, 1H), 3.95 (apparently s, 2H), 4.75 to 4.85 (m, 1H, partially overlapped) 5.10 (broad s, 1H) ppm.

^F3 C-NMR (125 MHz, D ₂ 0): guanidine δ 157.6; carbonyl carbons: δ 169.7, 170.2, 173.0.

(

Example 68

Method for producing MeOOC-CH ₂ ^- (R) Cha-Pic-Nag x 2 TFA

MeOOC-CH ₂ - (R) Cha-Pic-Nag (Z) methyl ester is obtained by transesterification of iprOOC-CH ₂ - (R) Cha-Pic-Nag (Z) (see Example 69) on a column during flash chromatography if eluent dichloromethane / methanol. The title compound is obtained by deprotection of method (a).

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.95-1.15 (m, 2H), 1.2-1.6 (m, 6H), 1.65-2.0 (m, 13H) 2.25 (broad d, 1H), 3.21 (t, 2H), 3.30 (t, 2H), 3.37 (m, 1H), 3.71 (m, 1H), 3, 83 (s, 3H), 3.97 (dd, 2H), 4.67 (broad t, 1H), 5.05 (broad s, 1H) ppm.

^P3 C-NMR (125 MHz, MeOD): guanidine: δ 158.0, carbonyl carbons: δ 173.0, 171.1 and 168.3 ppm.

Example 69

Method for producing &lt; RTI ID = 0.0 & gt _; PrOOC-CH2-(R) Cha-Pic-Nag &lt; / RTI &gt;

It is alkylated as in Example 4 using H- (R) ChaPic-Nag (Z) (see Example 6'5) and Βγ-ΟΗ ₂ ΟΟΟ ¹ Ργ followed by deprotection by method (a). The title compound is obtained.

¹ H-NMR spectrum (500 MHz, MeOD): δ 0.95-1.1 (m,

2H), 1.15-1.6 (m, 12H, of which 1.25 (d, 3H), 1.28 (d,

3H), 1.65-1.95 (m, 12H), 2.28 (broad d, 1H), 3.21 (t, 2H), 3.30 (t, 2H), 3.36 (m (1H), 3.93 (dd, 2H), 4.67 (t,

1H), 5.04 (broad s, 1H), 5.11 (pentet, 1H) ppm.

100 ¹³ C-NMR (125 MHz, MeOD): guanidine: δ 157.9, carbonyl carbons: δ 173.1, 171.0 and 168.3 ppm.

Example 70

Production method of HOOC-CH ₂ - (Me) - (R) Cha- (R or S) Pic-Nag / bx 2 TFA

It is alkylated as described in Example 4 using Me- (R) Cha- (R, S) Pic-Nag (Z) (see Example 66) and Br-CH ₂ COOBn followed by deprotection by method (b). HOOC-CH ₂ - (Me) - (R) Cha- (R or S) Pic-Nag is obtained. The two diastereomers were separated by reverse phase high performance liquid chromatography, eluting with 1: 3 acetonitrile: ammonium acetate, followed by removal of water and trifluoroacetic acid by lyophilization. This diastereomer is eluted as the last of the two diastereomers from the column.

1 H-NMR spectrum (500 MHz, MeOD): δ 0.9-1.1 (m,

2H), 1.15-1.35 (m, 4H), 1.4-1.55 (m, 2H), 1.6-1.85 (m, 12H), 2.3 (m, 1H) 2.85 (s, 3H), 3.15-3.45 (m, 5H), 3.65 (broad s, 2H), 4.0 (m, 1H), 4.65 (m, 1H) .5.08 (dd,

1H) ppm.

³³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.65, carbonyl carbons: δ 169.86 and 172.48 ppm.

Example 71

Production Method of HOOC- (R, S) CH (Me) - (R) Cha- (R, S) Pic-Nag x 2 TFA

Alkylation as in Example 4 using H- (R) Cha-Pic-Nag (Z) (see Example 65) and Br-CH (Me) -COOBn

101 followed by deprotection by method (a).

The title compound is obtained as a mixture of 4 diastereomers.

Example 72

Production method of HOOC- (R or S) CH (Me) - (R) Cha- (R or S) Pic-Nag / c x 2 TFA

The title compound is obtained by separating the diastereomers exemplified in Example 71 using reverse phase high performance liquid chromatography, eluting with a 1: 4 mixture of acetonitrile and 0.1 molar ammonium acetate, then evaporation and removal of water and trifluoroacetic acid by lyophilization. . This diastereomer was eluted as the third of four diastereomers from the column.

1 H-NMR spectrum (300 MHz, D ₂ O, 2 rotamers in approximately 5: 1 ratio): δ 0.88 (m, minor rotamer), 0.98 to 1.63 (m, 7H), 1.63 up to 2.02 (m, 16H, of which 1.68 (d, 3H)),

2.28 (m, 1H), 3.10 (t, minor rotamer), 3.25-3.50 (m,

5H, of which 3.33 (t, 2H) and 3.43 (t, 2H)), 3.82 (broad d,

1H), 4.02 (q, 1H), 4.55 (d, minor rotamer), 4.65 (d, minor rotamer), 4.72 (m, 1H), 5.10 (m, 1H) ppm .

Example 73

Production method of HOOC- (R or S) CH (Me) - (R) Cha- (R or S) Pic-Nag / d x 2 TFA

The title compound is obtained by separating the diastereomers exemplified in Example 71 using reverse phase high performance liquid chromatography, eluting with a 1: 4 mixture of acetonitrile and 0.1 molar ammonium acetate, then evaporation and removal of water and trifluoroacetic acid by lyophilization. This diastereomer elutes as the last of the four diastereomers from the column.

102 ^X H-NMR (500 MHz, D ₂ 0, 2 rotamers ca: 5: 1): δ 0.80 (m, minor rotamer), 0.90 (m, minor rotamer), 1.03 (m (2H), 1.10. to 1.33 (m, 3H), 1.42 (m, 2H), 1.51 to 1.92 (m, 16H, of which 1.57 (d, 3H)), 2.18 (d, 1H) ), 2.24 (d, minor rotamer), 2.98 (t, minor rotamer), 3.21 (t, 2H), 3.28-3.40 (m, 3H, thereof 3.44 (t (2H)), 3.82 (d, 1H), 4.02 (q, 1H), 4.42 (d, minor rotamer), 4.50 (t, minor rotamer), 4.62 (t, 1H) ), 4.67 (s, minor rotamer), 5.03 (s, 1H) ppm.

Example 74

Production method of HOOC-CH ₂ -CH ₂ - (R) Cha-Pic-Nag x 2 TFA

The compound was prepared from H- (R) Cha-Pic-Nag (Z) (see Example 65) in the same manner as described for HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Agm in Example 15 with the difference using 1.2 equivalents of benzyl acrylate instead of 1.1 equivalents.

¹ H-NMR spectrum (500 MHz, D ₂ 0, 2 rotamers in approximately 4: 1 ratio): δ 0.7 to 0.9 (m, minor rotamer), 0.9 to

1.0 (m, 2H), 1.05-1.3 (m, 3H), 1.3-1.45 (m, 2H), 1.5-

1.8 (m, 13H), 2.10 (d, 1H), 2.20 (d, minor rotamer),

2.75 (t, 2H), 2.95 (t, minor rotamer), 3.15 (t, 2H), 3.2-3.35 (m, 5H), 3.75 (d, 1H), 4.55 (t, 1H), 4.95 (m, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.57, carbonyl carbons: δ 170.16, 172.82 and 174.75 ppm.

Example 75

103

Production method of HOOC-CH ₂ - (R) Cha- (R, S) Mor-Agm x 2 TFA (i) Production method of H- (R) Cha-Mor-Agm (Z)

The compound was prepared from Boc- (R) Cha-Mor-OSu (see production of starting compounds) in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3.

(ii) Production method of HOOC-CH ₂ - (R) Cha- (R, S) Mor-Agm x 2 TFA

It is alkylated as in Example 4 using Br-CH ₂ COOBn followed by deprotection by method (b). The title compound is obtained. The epimerization of S-morphilin-2-carboxylic acid takes place to some extent during the synthesis and a mixture of two diastereomers, contained in a ratio of about 9: 1, is observed in the final substance.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.92-1.95 (m, 17H), 3.12-3.39 (m, 4H), 3.44-4.05 (m, 7H) ), 4.37 (d, 1H), 4.63 (m, 1H), 4.79 (broad d, 1H) ppm.

¹³ C-NMR (75.47 MHz, MeOD): guanidine: δ 158.63, carbonyl carbons: δ 170.87, 170.82 and 169.08, others δ 69.06 and 67.01 (COC) ppm .

Example 76

Production method of HOOC-CH ₂ - (R) Cha- (R or S) Mor-Nag x 2 TFA (i) Production method of H- (R) Cha-Mor-Nag (Z)

The compound is prepared from Boc- (R) Cha-Mor-OSu (see production of starting compounds) and Boc-Nag (Z) in the same manner as described for H- (R) Cha-Pro-Agm (Z) in Example 3 .

104 (ii) Production method of HOOC-CH ₂ - (R) Cha- (R or S) Mor-Nag x 2 TFA

It is alkylated as in Example 4 using Br-CH ₂ COOBn followed by deprotection by method (b). The title compound is obtained.

¹ H-NMR spectrum (300 MHz, MeOD): δ 0.92 to 1.13 (m,

2H), 1.15-1.42 (m, 3H), 1.50 (broad s, 1H), 1.62-1.95 (m, 9H), 3.14-3.40 (m, 4H) ), 3.46 to 4.13 (m, 7H), 4.41 (d, 1H), 4.63 (m, 1H), 4.80 (broad d, 1H) ppm.

¹³ C-NMR (75.47 MHz, MeOD): guanidine: δ 158.68, carbonyl carbons: δ 171.19, 170.90 and 169.46, others δ 68.81 and 67.00 (COC) ppm .

Example 77

Production method of H- (R) Cha-Aze-Nag x 2 HOAc (i) Production method of Boc- (R) Cha-Aze-Nag (Z)

The compound was prepared from Boc- (R) Cha-Aze-OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z) according to Example 65 (ic).

(ii) Production Method H. (R) Cha-Aze-Nag (Z)

The compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) (see Example 3).

(iii) Production method of H- (R) Cha-Aze-Nag x 2 HOAc

The compound is produced by applying method (a) of deprotection to product (ii). above.

105 ¹ H-NMR spectrum (300 MHz, D ₂ O): δ 0.85 to 1.10 (m,

2Η), 1.10 to 2.04 (m, 13H), 1.95 (s, acetate), 2.20 to 2.37 (m, 1H), 2.60 to 2.82 (m, 1H) 3.15 to 3.40 (m, 4Η), 3.96 to 4.15 (m, 2H), 4.18 to 4.30 (m, 1H), 4.30 to 4.42 (m, IH), minor rotamer signals are shown at δ 0.70, 3.90 and 5.10 ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.39, carbonyl carbons: δ 170.22 and 172.38 ppm.

Example 78

Production method of HOOC-CH ₂ - (R) Cha-Aze-Nag x HOAc (i) Production method of BnOOC-CH ₂ - (R) Cha-Aze-Nag (Z)

The compound was prepared from H- (R) Cha-Aze-Nag (Z) (see Example 77) according to the method described in Example 4.

(ii) _HOOC-CH2 - (R) Cha-Aze-Nag x HOAc

The compound is produced by applying method (a) of deprotection to the product (i) above.

1 H-NMR spectrum (500 MHz, MeOD): δ 0.90 to 1.10 (m,

2Η), 1.15 to 2.00 (m, (m, 1H), 2.58 to 2.70

13H '

1.95 (s, acetate), 2.20-2.30; m, 1H), 3.17-3.30 (m, 4H), 3.35-3.50 (m, 2H), 3, 55 to 3.68 (m, 1H), 4.10 to 4.20 (m, 1H), 4.30 to 4.38 (m, 1H), 4.65 to 4.77 (m, 1H), minor rotamer signals are shown at δ 3.75, 3.98, 4.03 and 5.08 ppm.

13,

C-NMR (75 MHz, D ₂ O): guanidine: δ 157.40, carbonyl carbons: δ 169.16, 171.92 'and 172.13 ppm.

ϊ

106

Example 79

Production method of H- (R) Cha-Pro (5- (S) Me) -Nag x 2 HCl (i) Production method of Boc- (R) Cha-Pro (5- (S) Me) -Nag (Z)

The same procedure used for the coupling of (R) Cha-OH and H-Pic-OEt x HCl (see production of starting compounds) was used to achieve coupling between Boc- (R) Cha-Pro (5- (S) Me) -OH and H-Nag (Z) x 2 HCl.

(ii) Production method of H- (R) Cha-Pro (5- (S) Me) -Nag (Z)

The same procedure as described for the synthesis of H- (R) -Cgl-Pic-Nag (Z) was used (see Example 84 (ii)).

(iii) A method for producing H- (R) Cha-Pro (5- (S) Me) -Nag x 2 HCl

The compound is produced by applying process (d) deprotection to the product (ii) obtained above.

¹ H-NMR spectrum (300 MHz, D ₂ O): δ 1.0-2.3 (m, 21H, thereof 1.47 (d, 3H)), 2.4-2.55 (m, 1H ), 3.3 to 3.6 (m,

4H), 4.30 (broad t, 1H), 4.38 (dd, 1H), 4.47 (broad t, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.6, carbonyl carbons: δ 174.6 and 169.6 ppm.

Example 80

Production method HOOC-CH ₂ - (R) Cha-Pro (5- (S) Me) -Nag x HOAc

Alkylation as in Example 4 using H- (R) Cha-Pro (5- (S) Me) -Nag (Z) (see Example 79) and the compound

107

Br-CH ₂ COOBn followed by deprotection by method (a). The title compound is obtained.

1 H-NMR spectrum (300 MHz, D ₂ O): δ 0.9-1.9 (m, 19H, of which 1.34 (broad d, 3H)), 1.93 (s, acetate), 2 0-2.2 (m, 3H), 2.34 (m, 1H), 3.1-3.5 (m, 7H), 3.97 (m, 1H), 4.20 (m, 1H) ), 4.31 (broad t, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.4 ppm.

Example 81

Method for producing HOOC-CHy (R) Cha- (R or S) Pie (4,5-dehydro) -Na / b X HOAC (i) Method for producing Boc- (R) Cha- (R, S) Pie (4, 5-dehydro) -Nag (Z)

This compound was prepared from Boc- (R) Cha- (R, S) Pie (4,5-dehydro) -OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z) (see example). 65 (ic)).

(ii) Production method of H- (R) Cha- (R, S) Pie (4,5-dehydro) -Nag (Z)

The title compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) (see example).

3).

(iii) Process for Producing Compound of _BnOOC-CH2 - (R) Cha- (R, S) Pie (4,5-dehydro) -Nag (Z)

The compound was prepared from H- (R) Cha- (R, S) Pie (4,5-dehydro) -Nag (Z) according to the method described in Example 4.

(iv) A method for producing the compound

108

HOOC-CH ₂ - (R) Cha- (R or S) Pie (4,5-dehydro) -Nag / bx HOAc

A mixture of 356 mg (0.539 mmol) of _BnOOC-CH2 - (R) Cha- (R, S) Pie- (4,5-dehydro) -Nag (Z), 10.8 ml of trifluoroacetic acid and 3.4 ml thioanisole Stir at room temperature for 3.5 hours. Water was added to the reaction mixture and the mixture was washed twice with dichloromethane and the solvent was evaporated. To give the compound _HOOC-CH2 - (R) Cha- (R, S) Pie (4,5-dehydro) -Nag. The title compound was obtained by separating the diastereomers by reverse-phase high performance liquid chromatography, eluting with a 3: 7 mixture of acetonitrile and 0.1 molar ammonium acetate, and then removing the water by lyophilization, after evaporation of the solvent. The diastereomer is eluted from the column as the second of the two diastereomers.

¹ H-NMR spectrum (300 MHz, D ₂ O): δ 0.85 to 1.95 (m, 15H), 2.50 to 2.80 (m, 2H), 3.25 (t, 2H), 3.35 (t, 2H), 3.55 (broad s, 2H), 3.85-4.6 (m, 3H), 4.92 (minor rotamer), 5.30 (d, 1H), 5 85 to 6.1 (m, 2H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.59, carbonyl carbons: δ 171.46, 172.58 and 173.03 ppm.

Example 82

Production method of HOOC-CH ₂ - (R) Cha-Pic (4- (S) Me) -Nag x 2 HCl (i) Production method of Boc- (R) Cha-Pic (4- (S) Me) -Nag ( OF)

The compound was prepared from Boc- (R) Cha-Pic (4- (S) Me) -OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z), according to the method of Example 65 (ic).

(ii) Production method of H- (R) Cha-Pic (4- (S) Me) -Nag (Z)

109

The title compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) (see Example 3).

(iii) BnOOC-CH ₂ - (R) Cha-Pic (4- (S) Me) -Nag (Z) production method

The compound was prepared from H- (R) Cha-Pic (4- (S) Me) -Nag (Z) according to the method described in Example 4.

(iv) Production method of HOOC-CH ₂ - (R) Cha-Pic (4- (R) Me) -Nag x 2 HCl

The compound is produced by applying process (d) deprotection to the product (iii) above.

1 H-NMR spectrum (500 MHz, D ₂ O): δ 0.95-2.05 (m, 22H, thereof 1.05 (d, 3H)), 2.30-2.38 (broad d, 1H), 3.28-3.36 (m, 2H), 3.36-3.50 (m, 3H), 3.85-3.95 (m, 1H),

3.98 (s, 2H), 4.70 to 4.90 (m, 1H, partially obscured by HOD signals), 5.22 to 5.27 (d, 1H), minor rotamer signal shown at δ 0.93 , 3.13 and 4.57 ppm.

³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.58, carbonyl carbons: δ 171.12, 170.32 and 172.82 ppm.

Example 83

Production method of HOOC-CH ₂ - (R) Cha- (R) Pic (4- (R) Me) -Nag x 2 HCl (i) Production method of Boc- (R) Cha- (R) Pie (4- (R) ) Me) -Nag (Z)

The compound was prepared from Boc- (R) Cha-Pic (4- (R) Me) -OS and Boc-Nag (Z) in the same manner as described for Boc- (R) Cha-Pro-Agm (Z) ( see example 3).

110 (ii) Production Method of H- (R) Cha- (R) Pie (4- (R) Me) -Nag (Z)

The title compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) (see example).

3).

(iii) _BnOOC-CH2 - (R) Cha- (R) Pie (4- (R) Me) -Nag (Z)

The compound was prepared from H- (R) Cha- (R) Pie (4- (R) Me) -Nag (Z) according to the method described in Example 4.

(iv) Production Method of HOOC-CH ₂ - (R) Cha- (R) Pie (4- (R) Me) -Nag x 2 HCl

The compound is produced by applying method (d) of deprotection to the product (iii) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 1.00 to 2.05 (m,

22H), 2.18-2.26 (broad d, 1H), 3.28-3.36 (m, 2H),

3.36 to 3.55 (m, 3H), 3.85 to 4.05 (m, 3H), 4.70 to 4.90 (m, 1H, partially overlapped by HOD signals), 5.35 to 5, 30 (d, 1H), minor rotamer signals appear at δ 2.40, 2.90,

4.10, 4.42, 4.55, and 5.23 ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 157.56, carbonyl carbons: δ 169.6.9, 169.84 and 173.20 ppm.

Example 84

Method for producing HOOC-CH ₂ - (R) Cgl-Pic-Nag x 2 HCl (i) Method for producing Boc- (R) Cgl-Pic-Nag (Z)

The compound was prepared from Bog- (R) Cgl-Pic-OH in the same manner as described above. is described for Boc- (R) Cha-Pic-Nag (Z), namely

111 according to the method of Example 65 (ic).

¹ H-NMR spectrum (300 MHz, CDCl 3): δ 0.9-1.8 (m, 27H), 2.4 (d, 1H), 3.1-3.3 (m, 5H), 3 9 (d, 1H), 4.2 (t, 1H), 5.1 (s, 2H), 5.2 (broad d, 2H), 6.7-7.4 (m, 9H) ppm.

(ii) Production Method of H- (R) Cgl-Pic-Nag (Z)

Hydrogen chloride gas was bubbled through a solution of 1.38 g (2.22 mmol) of Boc- (R) Cgl-Pic-Nag (Z) in 25 mL of ethyl acetate. After 10 minutes the solvent was evaporated and the residue was dissolved in ethyl acetate and 10% sodium carbonate solution. The organic phase was separated, washed with brine and dried over magnesium sulfate. Evaporation of the solvent gave 1.02 g of the title compound. Yield: 92%. @ 1 H-NMR spectrum (300 MHz, MeOD): .delta.1.0-1.9 (m, 18H), 2.2-2.3 (m, 1H), 3.2-3. 3 (m, 5H), 3.6 (d, 1 H),

3.8-3.9 (broad d, 1H), 4.2 (t, 1H), 4.7-4.8 (wide with 5H), 5.1 (s, 2H), 5.2 (s (1H), 7.2-7.3 (m, 5H) ppm.

(iii) BnOOC-CH ₂ - (R) Cgl-Pic-Nag (Z) production method

A solution of 291 mg (0.98 mmol) of trifluoromethanesulfonic acid ester with benzyl glycol in 2 ml of dichloromethane was added at -25 ° C to a stirred mixture of 0.52 g (1.04 mmol) of H- (R) Cgl-Pic-Nag ( Z) in 494 mg (3.58 mmol) of potassium carbonate in 5 mL of acetonitrile and 1 mL of dichloromethane. During the coupling, the temperature was allowed to rise to room temperature over 1 hour and after 5 days the reaction mixture was diluted with water and extracted with ethyl acetate and toluene. The organic phase is dried over magnesium sulphate and the solvent is evaporated. 319 mg of colorless crystals are obtained. Yield: 47%.

112 ¹ H-NMR spectrum (500 MHz,

1H), 1.1 to 1.3 (m, 4H), 1.35 to 8H), 1.8 to 2.2 (broad s, IH), 1H), 3.1 to 3.5 (m, 6H), 3.6 to 4H), 5.2 (with, IH) 6.5 to 7.4 (m,

CDClg): δ 1,0 to 1,1 (m,

1.6 (m, 5H), 1.6-1.85 (m,

2.23 to 2.5 (m, 2H), 2.9 (t,

3.7 (m, 2H), 5.0-5.1 (m,

13H) ppm.

(iv) A method for producing HOOC-CH ₂ - (R) Cgl-Pic-Nag x 2 HCl

319 mg (0.49 mmol) _BnOOC-CH2- (R) Cgl-Pic-Nag (Z) was dissolved with heating in 50 ml of isopropanol and 5 ml of water and hydrogenated over 228 mg of 10% palladium on carbon for 24 hours . After filtration and evaporation of the solvent followed by dissolution in hydrochloric acid, freeze-drying is performed. 223 mg of peptides were isolated as a white powder. Yield: 91%.

1 H-NMR spectrum (500 MHz, D ₂ O): δ 1.1-1.1 (m,

18H), 2.3 (d, 1H), 3.3 (t, 2H), 3.4 (t, 3H), 3.85-4.05 (m, 3H), 4.6 (d, 1H) 5.15 (s, 1H) ppm.

¹³ C-NMR (75 MHz, D ₂ O): guanidine: δ 157.43, carbonyl carbons: δ 169.2 and 172.94 ppm.

Example 85

Production Method of H- (R) Hoc-Pro-Nag x 2 TFA (i) Production Method of Boc- (R) Hoc-Pro-Nag (Z)

The compound was prepared from Boc- (R) Hoc-Pro-OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z) according to Example 65 (ic).

(ii) Production Method of H- (R) Hoc-Pro-Nag (Z)

113

The compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) (see Example 3).

(iii) Production method of H- (R) Hoc-Pro-Nag x TFA

The compound is produced by applying method (a) of deprotection to the product (ii) above.

¹ H-NMR spectrum (300 MHz, D ₂ O): δ 0.90-1.05 (m, 2H), 1.16-1.48 (m, 6H), 1.48-1.84 (m) 6H), 1.84-2.24 (m, 6H), 2.40 (m, 1H), 3.25-3.45 (m, 4H), 3.74 (m, 1H), 3, 85 (m, 1H); 4.42 (m, 1H); 4.51 (m, 1H) ppm.

Example 86

Production method of HOOC-CH ₂ - (R) Hoc-Pro-Nag x HOAc (i) Production method of BnOOC-CH ₂ - (R) Hoc-Pro-Nag (Z)

The compound was prepared from H- (R) Hoc-Pro-Nag (Z) (see Example 85) according to the method described in Example 4.

(ii) Production method of HOOC-CH ₂ - (R) Hoc-Pro-Nag x HOAc

The compound is produced by applying method (a) of deprotection to the product (i) above.

1 H-NMR spectrum (300 MHz, D ₂ O): δ 0.76 to 0.97 (m, 2H), 1.00 to 1.37 (m, 6H), 1.50 to 2.12 ( m, 12H), 1.89 (s, acetate), 2.27 (m, 1H), 3.10-3.33 (m, 4H), 3.41 (broad s 2H), 3.61 (m 1 H, 3.77 (m, 1 H), 4.12 (m, 1 H), 4.37 (m, 1 H) ppm.

C-NMR (75 MHz, D ₂ O): guanidine: δ 157.4, carbonyl carbons: δ 170.8, 173.9 and 174.5 ppm.

114

Example 87

Method for producing HOOC-CH ₂ - (R) Hoc-Pic-Nag x HOAc (i) Method for producing Boc- (R) Hoc-Pic-Nag (Z)

The compound was prepared from Boc- (R) Hoc-Pic-OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z) according to the method of Example 65 (ic).

(ii) Production method of H- (R) Hoc-Pic-Nag (Z)

The compound was prepared in the same manner as described for H- (R) Cha-Pro-Agm (Z) (see Example 3).

(iii) BnOOC-CH ₂ - (R) Hoc-Pic-Nag (Z) production method

The compound was prepared according to the method described in Example 4.

(iv) Production method of HOOC-CH ₂ - (R) Hoc-Pic-Nag x HOAc

The compound is produced by applying method (a) of deprotection to the product (iii) above.

1 H-NMR spectrum (300 MHz, D ₂ O): δ 0.75 to 0.95 (m, 2H), 1.00 to 1.30 (m, 6H), 1.30 to 1.50 (m (2H), 1.50 to 1.59

1.82 (m, 12H), 1.82-1.95 (wide s, acetate), 2.23 (wide d, 1H), 3.08-3.32 (m, 6H), 3.52 ( broad s, 2H), 3.77 (broad d, 1H), 4.50 (broad s, 1H), 5.00 (broad s, 1H) ppm

Example 88

Production method of HOOC-CH ₂ - (R) Dph-Pic-Nag x 2 HCl

115 (i) Manufacturing Method Boc- (R) Dph-Pic-Nag (Z)

The compound was prepared from Boc- (R) Dph-Pic-OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z) (according to Example 65 (ic)).

(ii) Production Method of H- (R) Dph-Pic-Nag (Z)

The compound was prepared in the same manner as described for H- (R) Cgl-Pic-Ag.m (Z) (see Example 84 (ii)).

(iii) BnOOC-CH ₂ - (R) Dph-Pic-Nag (Z) production method

The compound was prepared from H- (R) Dph-Pic-Nag (Z) according to the method described in Example 4.

(iv) A method for producing HOOC-CH ₂ - (R) Dph-Pic-Nag x 2 HCl

The compound is produced by applying process (d) of deprotection to the product (iii) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 0.46 (m, 1H), 1.2-1.35 (m, 2H), 1.45 (m, IH), 1.53 ( m, 1H), 1.89 (pentet, 2H), 2.03 (broad d, 1H), 3.24 (broad t, 1H), 3.29 (t, 2H), 3.38 (t, 2H), 3.72 (d, 1H), 3.78 (d, IH), 3.79 (m, IH), 4.68 (d, IH), 4.89 (m, IH), 5, 73 (d, 1H), 7.4-7.6 (m, 6H), 7.65 (t,

2H), 7.81 (d, 2H) ppm.

Example 89

HOOC-CH ₂ ^_ (R) Dch-Pic-Nag x HOAc (i) Boc- (R) Dch-Pic-Nag (Z)

116

The compound was prepared from Boc- (R) Dch-Pic-OH in the same manner as described for Boc- (R) Cha-Pic-Nag (Z) (according to Example 65 (ic)).

(ii) Production Method of H- (R) Dch-Pic-Nag (Z)

The compound was prepared in the same manner as described for H- (R) Cgl-Pic-Nag (Z) (see Example 84 (ii)).

(iii) BnOOC-CH ₂ - (R) Dch-Pic-Nag (Z) production method

The compound was prepared from H- (R) Dch-Pic-Nag (Z) according to the method described in Example 4.

(iv) _HOOC-CH2 - (R) Dch-Pic-Nag x HOAc

The compound is produced by applying method (a) of deprotection to the product (iii) above.

¹ H-NMR spectrum (500 MHz, D ₂ O): δ 1.2 to 2.0 (m,

30H), 2.09 (s, acetate), 2.30 (broad d, 1H), 3.32 (t, 2H), 3.4 to 3.5 (m, 3H), 3.65 (d, 1H, 3.70 (d, 1H), 3.86 (broad d, 1H), 4.86 (m, 1H), 5.09 (m, 1H) ppm.

¹³ C-NMR (125 MHz, D ₂ O): guanidine: δ 159.4, carbonyl carbons: δ 172.5, 173.3 and 174.9 ppm.

Example PI

Solution for parenteral administration

The solution is prepared from the following components:

HOOC-CH ₂ - (R) Cha-Pic-Nag x 2 HBr sodium chloride for injection

Acetic acid 3 g water for injections up to 1000 ml

The active ingredient, sodium chloride and acetic acid are dissolved in water. The pH is adjusted to 3 to 7 with 2M sodium hydroxide solution. The solution is filtered through a sterile 0.2 µm filter and filled into sterile ampoules under aseptic conditions.

Example P2

Tablets for oral administration

1000 tablets are prepared from the following ingredients:

thrombin inhibitor 100 g lactose 200 g polyvinylpyrrolidone 30 g microcrystalline cellulose 30 g magnesium stearate 6 g

The active ingredient and lactose are mixed with an aqueous solution of polyvinylpyrrolidone. The mixture is dried and milled to form granules and then microcrystalline cellulose is added to the granules followed by magnesium stearate. The mixture is then compressed on a tabletting machine to give 1000 tablets, each containing 100 mg of active ingredient.

Biological tests

Determination of clotting time of thrombin and IC ₅₀ TT

Human thrombin (T 6769, Sigma Chem. Co.) is incubated for 1 minute in 100 μ 100 buffer solution pH 7.4 and 100 μΐ inhibitor solution. The solution is then added

118 adds 100 μΐ of citrate-treated normal human plasma and the clotting time is measured on an automatic KC 10 instrument (Amelung).

The clotting time, expressed in seconds, is plotted against inhibitor concentration and IC ₅₀ TT values are determined by interpolation.

IC ₅₀ TT represents the concentration of inhibitor that results in twice the clotting time of thrombin for human plasma. pIC ₅₀ TT is -log 10 of IC ₅₀ TT in mol / _L. Preferred compounds of the invention have pIC ₅₀ TT values ranging from 6.6 to 8.2.

Determination of Thromboplastin Partial Activation Time (APTT)

APTT is determined in a citrate-treated normal human plasma stock that has been contacted with PTT Automated 5 reagent manufactured by Stago. Plasma (10 μΐ inhibitor solution per 90 μΐ plasma) is added to the inhibitors and APTT is measured in the mixture using a KC 10 coagulation analyzer (Amelung) according to the reagent manufacturer's instructions. The clotting time, expressed in seconds, is plotted against the inhibitor concentration in plasma and IC ₅₀ APTT values are determined by interpolation.

IC _5Q APTT is defined as the concentration of inhibitor in plasma that doubles the Activated partial thromboplastin time. pIC _5Q APTT is -log 10 of IC _5Q APTT in mol / _L. Among the compounds, the compounds of the present invention are preferred in which the tests show ρΙΟΙΟ θΑΡΤΤ values ranging from 5.1 to 6.4.

119

Overview of commonly used abbreviations

Agm = Agm (Z) = AA ₁ = a and ₂ = Aze = agmatin oo-N-benzyloxycarbonylagmatin amino acid 1 amino acid 2 (S) -azetidine-2-carboxylic acid Bia = α-substituted butyrolactone Boc ~ tert-butoxycarbonyl Bu = butyl Bn = benzyl Cgl = Ch = (S) -cyclohexylglycine cyclohexyl Cha = (S) -β-Cyclohexylalanine

CME-CDI = 1-cyclohexyl-3- (2-morpho-metho-p-toluenesulfonate)

(ethylethyl) carbodiimide DCC = dicyclohexylcarbodiimide Dch = (S) -dicyclohexylalanine DMAP = N, N-dimethylaminopyridine DMF = dimethylformamide DMSO = dimethylsulfoxide Dph = EDC = (S) -diphenylalanine 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride Et - ethyl EtOAc = ethyl acetate HOAc = acetic acid HOBt = N-hydroxybenzotriazole Hoc = (S) -homocyclohexylalanine Hop = HOSU = (S) -homophenylalanine N-hydroxysuccinimide Mag = Me = miniagmatin methyl Mor = (S) -morpholine-2-carboxylic acid

120

MPa = pressure expressed in 10 ⁶ pascal Nag = noragmatin Nag (Z) = δ-N-benzyloxycarbonylnoragmatine NMM = N-methylmorpholine Pgl = (S) -phenylglycine Ph = phenyl Phe = (S) -phenylalanine Pie - (S) -pipecolinic acid Pr = propyl Pro = (S) -proline RPLC = high performance liquid chromatography versní phase Tf = trifluoromethylsulfones1 TFA = trifluoroacetic acid THF = tetrahydrofuran p-TsOH = p-toluenesulfonic acid Val = (S) -valine Z = benzyloxycarbonyl

The refixes n, s, i and t, respectively η-, sec-, iso and t-have their usual meanings and mean normal secondary and tertiary.

iso

Scheme I (Examples 3 to 18, 20 and 21, 24 to 28, 30 to 34, 36 to 40, 43 to 49, 51 to 53, 57 to 64, and 67 to 93)

Boc-AA- ₁ -AA ₂ -OH coupling with H-NAG (Z) or H-Agm (Z)

Boc-AA ₁ -AA ₂ -Nag (Z) (or Agm (Z))

1. removing the protecting group from the terminal nitrogen

2. Reaction of terminal nitrogen with electrophilic compound (see special examples for detailed information)

AA ₁ -AA ₂ ~ Nag (Z) (or Agm (Z)) removal of the protecting group or groups

AA ^ -AA ₂ ~ Nag (or Agm)

AA _T = H- (R) Cha-OH; Me- (R) Cha-OH; H- (R, S) Pro (3- (trans) Ph-OH; H- (R) Hoc-OH; (R) Cgl-OH, H- (R) Dph-OH, H- (R) Dch-OH

AA ₂ = H-Pro-OH, H-Pic-OH, H-Mor-OH, H-Aze-OH, H-Pic (4-S) -OH, H-Pic (4- (R)) Me) -OH

H- (R, S) Pc (4,5-dehydro) -OH, H- (R) Pie (4- (R) Me) -OH, H-Pro (5- (R, S) Me) -OH H-Pro (5- (S) Me) -OH

H-Pic (6- (S) Me) -OH

122

N-terminal group in the final compound =

H, HO- (CH _{2) 3} -, ⁿ Bu-, HOOC-CH2-, MeOOC-CH2-, ⁱ PrOOC-CH2, ^t BuOOC-CH ₂ -, HOOC-CH (Me) -,

HOOC-CH ( ⁿ Pr) -, HOOC-CH (Ph) -, HOOC-CH (CH ₂ CH ₂ Ph) -, HOOC-CH ₂ CH ₂ -, HOOC-CH ₂ CH ₂ CH ₂ -, EtOOC-CH ₂ CH ₂ CH ₂ -, Bia, HOOC-CH ₂ -OOC-CH ₂ -, EtOOC-CO-, MeOOC-CO-, HOOC-CO-, H ₂ NOC-CH ₂ -, HOOC-CH (CH ₂ COOH ) -, MeOOC-CH (CH ₂ COOMe) -, HOOC-CH ₂ -NH-CO-CH ₂ -, HOOC-CH (CH ₂ OH) -, (HO) ₂ P (O) -CH ₂ -, EtO (HO) P (O) -CH ₂ -, (EtO) ₂ P (O) -CH ₂ -.

Scheme II (Examples 55, 56, 65 and 66)

Boc - (R, S) Pro (3-Ph) -OH

H-Pro-OBn, HOBt, NMM, DMF

The second H _2, Pd / C

Third HOSu, CME-CDI, _CH3 CN

H-Agm (Z);

Boc- (R, S) Pro (3-Ph) Pro-OSu -> Boc- (R, S) Pro (3-Ph) Pro-Agm (Z)

DMF, temp. rooms

First TFA, CH ₂ C1 ₂

The second H _2, Pd / C

H- (R, S) Pro (3-Ph) -Pro-Agm Example 55

Rh / Al ₂ O ₃ , H ₂

Boc- (R, S) Pro (3- (trans) Ph) -OH → Boc- (R, S) Pro (3- (trans) Ch) -OH HOAc, MeOH see Example 55 (above)

Boc- (R) Cha-OH H- (R, S) Pro (3- (trans) Ch) -Pro-Agm Example 56

First HOBt, CME-CDI, CH ₂ C1 ₂

2. HCl x H-Pic-OEt, NMM, DMF

3. LiOH, THF, H ₂ O ^1f 4. HOS, DCC, DMF

Boc - (R) Cha - Pic - OS Boc - (R) Cha - Pic - Nag (Z)

H-Nag (Z), NMM or -> or

DMF, temp. rooms

Boc- (Me) (R) Cha- (R, S) Pic-OS Boc- (Me) (R) Cha- (R, S) Pic-Nag (Z)

124 η 1. HOBt, CME-CDI, CH ₂ Cl ₂

2. HCl x H-Pic-OEt, NMM, DMF

3. LiOH, THF, H ₂ O

4. HOS, DCC, DMF H - (R) Cha - Pic - Nag

Boc- (Me) (R) Cha-OH or. TFA second H _2, Pd / C (Example 65) (Me) (R) Cha- (R, S) Pic-Nag Example 66

Scheme III (Examples 1 and 2)

H-AgmxHCl, Et ₃ N

Boc (R) Cha-Pro-OSu -> Boc (R) Cha-Pro-Agm

DMF, temp. rooms

TFA

H- (R) Cha-Pro-Agm Example 1

H-AgmxHCl, NMM

Boc- (Me) Cha-Pro-OSu -> Boc- (Me) Cha-Pro-Agm

DMF, temp. rooms I

I TFA

Me- (R) Cha-Pro-Agm Example 2

126

Scheme IV (Example 19)

H ₂ N- (CH ₂ ) ₃ -NH (Boc)

Z- (R) Cha-Pro-OSu -> Z- (R) Cha-Pro-NH- _{(CH2) 3} -NH (Boc)

NMM, DMF, Temp. rooms

TFA ’r

Z- (R) Cha-Pro-NH- (CH _{2) 3} -NH ₂

1. 3,5'-dimethyl-1-pyrazolylformamidinium nitrate, Et ₃ N, DMF ,, 2. H ₂ , Pd / C

H- (R) Cha-Pro-Nag Example 19

X 4. i

Scheme V (Example 54)

Boc (R) Cha-Pro-OS

H ₂ N- (CH ₂ ) ₂ -NH (Z),

NMM, DMF, Temp. rooms

Boc- (R) Cha-Pro-NH- _{(CH2) 2-NH} (Z). TFA

2. BrCH ₂ COOEt, CH ₃ CN, K ₂ CC> ₃

EtOOC-CH ₂ - (R) Cha-Pro-NH- (CH _{2) 2} -NH (Z)

The first H _2, Pd / C

2. 3,5'-Dimethyl-1-pyrazolylformamidinium nitrate, Et-N, DMF

3. NaOH / EtOH

EtOOC-CH ₂ - (R) Cha-Pro-Mag Example 54

128

Scheme VI (Examples 22, 23, 29, 35, 41, 42 and 50)

_HOOC-CH2 - (R) Cha-Pro-Nag Example 22

NaOH / EtOH

EtOOC-CH ₂ - (R) Cha-Pro-Nag Example 23 (HOOC-ch ₂ ) ₂ - (R) Cha-Pro-Nag Example 29

NaOH / EtOH (EtOOC-CH ₂ ) ₂ - (R) Cha-Pro-Nag

1. BrCH ₂ COOEt, CH ₃ CN, K ₂ CO ₃

The second H _2, Pd / C

3. 3,5'-dimethyl-1-pyrazolylformamidinium nitrate,

Et ₃ N, DMF

1. BrCH ₂ COOEt (excess), K ₂ CO ch ₃ cn

The second H _2, Pd / C

3. 3,5'-Dimethyl-1-pyrazolylformamidinium nitrate, Et ₃ N, DMF

Boc (R) Cha-Pro-OS

1. H ₂ N- (CH ₂ ) ₃ -n ₃ NMM, DMF

-> H- (R) Cha-Pro-NH- (CH ₂ ) ₃ -N ₃

-pyrazolylformamide inium nitrate, Et ₃ N, DMF

First = _CH2 COOEt, EtOH

The second H _2, Pd / C

3. 3,5-dimethyl-1-pyrazolylformamide inium nitrate, Et ₃ N, DMF

EtOOC- (R, S) CH (Me) - (R) Cha-Pro-Nag EtOOCCH ₂ CH ₂ - (R) Cha-Pro-Nag Example 35 Example 42 izy

NaOH / EtOH

H- (R) Cha-Pro-NH- (CH ₂ ) ₃ -N ₃ HOOCCH ₂ CH ₂ - (R) Cha-Pro-Nag Example 41

1. ^ΕΒΒΟΟΟ -η1η-4-ΟΗ ₂ ΒΓ, K ₂ C ° ₃ , CH ₃ CN

2. bis-phenylthiostannane, PhSH, Et ₃ N

3. 3,5-dimethyl-1-pyrazolyl-formamidinium nitrate, Et ₃ N, DMF

TFA ^ΕυΒ -Ρίι-4-ΟΗ _2- (R) Cha-Pro-Nag -> HOOC-Ph-4-CH ₂ ~ (R) Cha-Pro-Nag Example 50 (original version)

¹³⁰ i 3, ir!

ί i

AND

Claims (1)

PATENT i_N changed based on > rt o! O '-O z L / i X O O i ci czx OC · O ; · .X i R 0 K Y ~ Xi) of the day 8.7.1994) A compound of formula (CH 3 _n -B wherein A is methylene or A represents an ethylene group and the resulting five-membered ring may or may not contain one or two fluorine, hydroxy or oxo at the 4-position, or may or may not be saturated, or A is -CH ₂ -O-, -CH ₂ -S- or -CH ₂ -SO- with a heteroatom functionality at the 4-position, or A represents an n-propylene group and the resulting six-membered ring may or may not contain one fluorine, hydroxy or oxo group at the 5-position, two fluorine atoms at one of the 4 or 5 positions, or unsaturated at the 4 and 5 positions or (C 1 -C 4); or A is -CH ₂ -O-CH ₂ -, -CH ₂ -S-CH ₂ ( 131 or -CH ₂ -SO-CH ₂ -, R ³ represents a hydrogen atom, a C 1 -C 4 alkyl group, a C 2 or C 3 hydroxyalkyl group or a group of formula R ³³ OOC-alkyl wherein the alkyl group contains 1 to 4 carbon atoms and R ³³ represents a hydrogen atom or (C 1 -C 4) -alkyl or (C 2 -C 3) -alkylene α intramolecularly bound to the carbonyl group at R ³ ; or R ³ represents R ³² OOC-1,4-phenyl-CH ₂ -, wherein R ³² represents a hydrogen atom or a C 1 -C 4 alkyl group, or R ³ represents a group of formula R ³³ -NH-CO-alkyl-, wherein alkyl as the alkyl group contains 1 to 4 carbon atoms and is optionally substituted at the α position to the carbonyl group with a C 1 -C 4 alkyl group and wherein R ³³ represents an atom hydrogen or alkyl having 1 to 4 carbon atoms or represents a group of formula -CH ₂ -COOR ^32, wherein R ³² is as defined above, or R ³ represents a group of the formula R ³² OOC-CH ₂ -OOC-alkyl-, wherein alkyl as the alkyl group contains 1 to 4 carbon atoms and is optionally substituted at the α position to the carbonyl group with a C 1 -C 4 alkyl group and wherein R has as defined above, or R ³ is CH ₃ SO ₂ -, or R ³ represents a group of formula R ³² OCOCO-, wherein R ³² is as defined above, or R ³ is -CH ₂ PO (OK ³⁴ ) 2, -CH ₂ SO ₃ H 132 or -CH ₂ - (5 (1 H) -tetrazolyl), wherein R ² - ⁴ is each independently hydrogen, methyl or ethyl, R ² represents a hydrogen atom, a C 1 -C 4 alkyl group or a group of the formula R ²¹ OOC-alkyl-, wherein the alkyl group contains from 1 to 4 carbon atoms and is optionally substituted in position a to the carbonyl group and a substituent in position a is a group of formula R ²² - (CH ₂ ) p - wherein p is 0 to 2 and R ²² is methyl, phenyl, hydroxy or a group of formula COOR ²² · and wherein R ²² · represents a hydrogen atom or an alkyl group of 1 to 2; 4 carbon atoms, m is 0, 1 or 2, R ³ represents a cyclohexyl group and R ⁴ represents a hydrogen atom, or m is 1, O R is cyclohexyl or phenyl; and R ⁴ forms an ethylene bridge together with R ¹ , or m represents the number 1 a R ³ and R ⁴ are each cyclohexyl or phenyl, R ⁵ represents a hydrogen atom or a C 1 -C 4 alkyl group, n represents an integer from 2 to 6 a B represents a group of the formula -N (R ⁶ ) -C (NH) -NH ₂ wherein R ⁶ represents a hydrogen atom or a methyl group, or B is -SC (NH) -NH ₂ or -C (NH) -NH ₉ , 133 wherein the compound is either as such or in the form of a physiologically acceptable salt and comprising isomers. A compound according to claim 1, wherein R ¹ represents a group of formula R ¹¹ OOC-alkyl-, wherein the alkyl group contains from 1 to 4 carbon atoms and R ¹¹ represents a hydrogen atom. A compound according to claim 2, wherein A represents ethylene and R ⁵ represents a hydrogen atom or a C 1 -C 4 alkyl group. The compound of claim 2, wherein A is n-propylene and the resulting six-membered ring may or may not have a C 1 -C 4 alkyl group at the 4-position and R is hydrogen or C 1 -C 4 alkyl. A compound according to at least one of the preceding claims, wherein R ³ represents a cyclohexyl group, m represents a number 1 or 2 and R ⁴ represents a hydrogen atom. A compound according to at least one of the preceding claims, wherein n is the number 3. A compound according to at least one of the preceding claims having an S-configuration at the α-amino acid position A compound according to claim 7 having the R-configuration at the α-amino acid at the P ₃ position. A compound selected from the group consisting of H- (R) Cha-Pro-Agm αúžúžttúžttis í; íá + Wteíatto ·; 134 Me (R) Cha-Pro-Agm HO- (CH _{2) 3} - (R) Cha-Pro-Agm ⁱ _PrOOC-CH2 - (R) Cha-Pro-Agm HOOC - (R, S) CH (Me) - (R) Cha-Pro-Agm HOOC- (R or S) CH (Me) - (R) Cha-Pro-Agm / and HOOC- (R or S) CH ( ⁿ Pr) - (R) Cha-Pro-Agm / and HOOC- (R or S) CH ⁽ⁿ Pr) - (R) Cha-Pro-Agm / b HOOC- (R) CH (Ph) - (R) Cha-Pro-Agm / b HOOC- (R, S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm HOOC- (R or S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm / a HOOC-CH ₂ -CH ₂ - (R ) Cha-Pro-Agm EtOOC-CO - (R) Cha - Pro - Agm (R, S) - Bia - (R) Cha - Pro - Agm HOOC- (R or S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Agm / b H- (R) Cha-Pro-Nag ⁿ Bu- (R) Cha-Pro-Nag HO- (CH ₂ ) 3 - (R) Cha-Pro-Nag EtOOC-CH ₂ - (R) Cha-Pro-Nag ⁱ _PrOOC-CH2 - (R) Cha-Pro-Nag ^t _BuOOC-CH2 - (R) Cha-Pro-Nag HOOC-CH ₂ -OOC-CH ₂ - (R) Cha-Pro-Nag H ₂ N-CO-CH ₂ - (R) Cha-Pro-Nag HOOC-CH ₂ -NH-CO-CH ₂ - (R) Cha-Pro-Nag (HOOC-CH ₂ ) _2- (R) Cha-Pro-Nag HOOC-CH. Bu) - (R) Cha-Pro-Nag HOOC - (R, S) CH (Me) - (R) Cha - Pro - Nag HOOC - (R or S) CH (Me) - (R) Cha - Pro - Nag and EtOOC - (R, S) CH (Me) - (R) Cha-Pro-Nag HOOC- (R) CH ⁽ⁿ Pr) - (R) Cha-Pro-Nag / a HOOC- (R) CH (CH ₂ OH) - (R ) Cha-Pro-Nag HOOC - (R, S) CH (Ph) - (R) Cha-Pro-Nag HOOC- (S) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Nag HOOC- (R) CH (CH ₂ CH ₂ Ph) - (R) Cha-Pro-Nag HOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Nag EtOOC-CH ₂ -CH ₂ - (R) Cha-Pro-Nag 135 HOOC- (CH ₂ ) 3 - (R) Cha-Pro-Nag EtOOC- (CH ₂ ) ₃ - (R) Cha-Pro-Nag HOOC-CO - (R) Cha - Pro - Nag MeOOC-CO - (R) Cha - Pro - Nag (R, S) Bia - (R) Cha - Pro - Nag HOOC- (R, S) CH (CH ₂ COOH) - (R) Cha-Pro-Nag MeOOC- (R, S) CH (CH ₂ COOMe) - (R) Cha-Pro-Nag _{HOOC-Ph-4-CH2} - (R) Cha-Pro-Nag (HO) ₂ P (0) _-CH2 - (R) Cha-Pro-Nag EtO (HO) P (O) -CH ₂ - ( R) Cha-Pro-Nag (EtO) ₂ P (0) _-CH2 - (R) Cha-Pro-Nag _HOOC-CH2 - (R) Cha-Pro-Mag H- (R, S) Pro (3 Ph) -Pro-Agm H- (R, S) Pro (3- (trans) Ch) -Pro-Agm _HOOC-CH2 - (R, S) Pro (3- (trans) Ph) -Pro-Agm HOOC-CH ₂ - (R, S) Pro (3- (trans) Ph) -Pro-Nag HOOC-CH ₂ (Me) - (R) Cha- (R, S) Pic-Agm ) CH (Me) - (R) Cha-Pic-Agm HOOC- (R or S) CH (Me) - (R) Cha-Pic-Agm / a _{HOOC-CH2 -CH2} - (R) Cha-Pic -Agm H- (R) Cha-Pic-Nag Me- (R) Cha- (R, S) -Pic-Nag MeOOC-CH ₂ - (R) Cha-Pic-Nag ^and PrOOC-CH ₂ - (R) Cha-Pic-Nag HOOC-CH ₂ - (Me) (R) Cha- (R) Pic-Nag / b HOOC- (R, S) CH- (Me) - (R) Cha- (R, S.) Pic-Nag HOOC- (R or S) CH (Me) - (R) Cha- (R or S) Pic-Nag / c _{HOOC-CH2 -CH2} - (R) Cha-Pic-Nag _HOOC-CH2 - (R) Cha- (R, S) Mor-Agm _HOOC-CH2 - (R) Cha- (R or S) Mor-Nag H - (R) Cha-Aze-Nag _HOOC-CH2 - (R) Cha-Aze-Nag H- (R) Cha-Pro (5- (S) Me) -Nag HOOC-CH ₂ - (R) Cha- (R or S) Pie (4, 5-dehydro) -Nag / b HOOC-CH ₂ - (R) Cha- (R) Pic (4- (R) Me) - Nag 136 _HOOC-CH2 - (R) Cgl-Pic-Nag H- (R) Hoc-Pro-Nag _HOOC-CH2 - (R) Hoc-Pro-Nag _HOOC-CH2 - (R) Hoc-Pic-Nag _HOOC-CH2 - (R) Dph-Pic-Nag _HOOC-CH2 - (R) Dch-Pic-Nag _HOOC-CH2 - (R) Cha-Pro (5- (R, S) Me) -Nag _HOOC-CH2 - (R) Cha-Pic (4- (R) Me) -Nag H- (R) Cha -Pic (4- (R) Me) -Nag HOOC-CH ₂ - (R) Cha-Pic (6- (S) Me) -Nag, wherein the compound is either as such or in the form of a physiologically acceptable salt and comprising isomers . A compound selected from the group consisting of _HOOC-CH2 - (R) Cha-Pro-Agm HOOC-CH ₂ - (Me) - (R) Cha-Pro-Agm HOOC- (R or S) CH (Me) - (R) Cha-Pro-Agm / b _HOOC-CH2 - (R) Cha-Pro-Nag _HOOC-CH2 - (R) Cha-Pic-Agm HOOC - (R or S) CH (Me) - (R) Cha-Pic-Agm / b HOOC- (R or S) CH (Me) - (R) Cha- (R or S) Pic-Nag / d _HOOC-CH2 - (R) Cha-Pro (5- (S) Me) -Nag HOOC-CH ₂ - (R) Cha-Pic (4- (S) Me) -Nag wherein the compound is either as such or in the form of a physiologically acceptable salt and comprising isomers. Compound H00C-CH ₂ - (Me) - (R) Cha-Pro-Nag, either as such or in the form of a physiologically acceptable salt and comprising isomers. The compound HOOC- (R or · S) CH (Me) - (R) Cha-Pro-Nag / b, either as such or in the form of a physiologically acceptable salt C. ·,. ·. 137 and including isomers. The compound HOOC-CH ₂ - (R) Cha-Pic-Nag, either as such or in the form of a physiologically acceptable salt and comprising isomers. 14. A process for producing a compound according to any of claims 1 to 13, characterized in that the coupling of an N-terminally protected amino acid or dipeptide or a preformed N-terminal alkylated protected dipeptide to a compound ^H 2 N- (C ^H 2) _n X wherein n is an integer from 2 to 6 a X is an unprotected or protected guanidino, protected amino or convertible amino group, which is subsequently converted to a guanidino group and, if desired, a physiologically acceptable salt is formed, and in such cases where the reaction results in a mixture of stereomers these stereomers are optionally separated by standard chromatographic or recrystallization techniques, and if desired, a single stereomer is isolated. Process according to claim 14, for the preparation of a compound according to any one of claims 1 to 13, characterized in that: a) (method I), a N-terminally protected dipeptide is coupled with either protected or unprotected guanidine or alkylamine 138 straight chain, protecting or masking the amino group at the end of the alkyl chain, using the standard peptide coupling shown in the formula H ₂ N- (CH ₂ ) _n -X in which formulas 4 5. R of R, R, n, m and A have the meanings given in the general formula AND, R ^b represents a hydrogen atom or an alkyl group, an amino protecting group such as tert-butoxycarbonyl and benzyloxycarbonyl; and 139 represents a group of formula -NH-C (NH) NH ₂ , -NH-C (NH) NH-W ₂ , -N (W ₂ ) -C (NH) NH-W ₂ , -NH-C (NW ₂ ) NH-W ₂ or -NH-W ₂ , wherein W ₂ represents an amino protecting group such as tert-butoxycarbonyl and benzyloxycarbonyl, or represents a masked amino group such as an azide residue, to form a protected peptide, or b) (method II), the N-terminal protected amino acid, with either protected or unprotected aminoguanidine or straight chain alkylamine, is coupled to protect or mask the amino group at the end of the alkyl chain, using the standard peptide coupling shown in the formula W, —N A OH Η ₂ Ν- (ΟΗ2) _η -Χ 140 RW-N A ⁰ NH- (CH ₂ ) _n -X in which formulas W ₃ , A, R ³ _and x have the meanings given above, followed by removal of the W ₂ group and coupling with the N-terminal amino acid in protected form, or c) (method III), a preformed N-terminal alkylated or protected dipeptide, produced by standard peptide coupling, is coupled with either protected or unprotected aminoguanidine or straight chain alkylamine, while protecting or masking the amino group at the end of the alkyl chain, using a standard peptide the coupling shown by the formula H ₂ N- (CH ₂ ) _n -X Wr R ² ' R ³ X N ‘ R '(C HR 1 NA ⁰ NH- (CH ₂ ) _n -X in which formulas R ² , R ³ , R ⁴ , R ⁵ , n, m, A and X have the meanings given above, provided that R ² is other than hydrogen, and W ₃ represents an acyl protecting group such as a trifluoroacyl group, whereupon the final compounds are prepared by any of the following methods, depending on the nature of the X- group used, by removing the protecting group or protecting groups when X- is -NH-C (NH) NH _2, -NH-C (NH) NH-W ₂ , -N (W ₂ ) -C (NH) NH-W ₂ or -NH-C (NW ₂ ) NH-W ₂ , or by selective removal of the protecting group W -? - for example when X represents a group of formula -NH-C (NH) NH-W _2, -N (W ₂ ) -C (NH) NH-W ₂ or -NH-C (NW ₂ ) NH-W ₂ , where W ₂ in this case must be placed orthogonal to, followed by alkylation of the N-terminal nitrogen and removal or by selective deprotection or unmasking of the terminal alkylamino function when X is -NH-W ₂ , where W ₂ in this case must be placed orthogonal to W a W ₃ , or X is a masked amino group such as an azide moiety, followed by guanidization reaction of the free amine, using standard methods, and removing the group W 1 - or 142 and, if desired, a physiologically acceptable salt is prepared, and in those cases where the reaction results in a mixture of stereomers, these stereomers are optionally separated by standard chromatographic or recrystallization techniques, and if desired, a single stereomer is isolated. Wo-, Use of a compound of formula NH NH ₂ either as such or in the form of a salt and as such or containing guanidino, either protected once on δ-nitrogen or twice protected on δ-nitrous or τ, δ-nitrous, as starting compounds for the synthesis of a serine protease inhibitor and in particular for synthesis thrombin inhibitor. Use according to claim 16, characterized in that the serine protease inhibitor is a peptide compound. 18. Structural fragment of the formula NH O as a structural element of a pharmaceutically active compound. 19. The structural fragment of claim 18, wherein the pharmaceutically active compound is a peptide compound. J A compound according to any one of claims 1 to 13 for use in therapy. The compound of claim 20 for use as an anticoagulant and antithrombotic agent. 22. A pharmaceutical composition comprising an effective amount of a compound of claims 1 to 13 together with at least one pharmaceutical carrier. The pharmaceutical composition of claim 22 for use as an anticoagulant and antithrombotic agent. Use of a compound according to any one of claims 1 to 13 as an active ingredient for the manufacture of a pharmaceutical composition for inhibiting thrombin in a human or animal body. A compound, method, pharmaceutical composition, use and method of use according to any one of claims 1 to 24 and substantially as described.